Squaric acid derivatives

ABSTRACT

Squaric acid Derivatives of formula (1) are described:  
                 
 
     wherein  
     R 1  is an integrin binding group;  
     R 2  is a hydrogen atom or a C 1-6 alkyl group;  
     L 1  is a covalent bond or a linker atom or group;  
     n is zero or the integer 1;  
     Alk 1  is an optionally substituted aliphatic chain;  
     R 3  is a hydrogen atom or an optionally substituted heteroaliphatic, cycloaliphatic, heterocycloaliphatic, polycycloaliphatic, polyheterocycloaliphatic, aromatic or heteroaromatic group:  
     and the salts, solvates, hydrates and N-oxides thereof.  
     The compounds are able to inhibit the binding of integrins to their ligands and are of use in the prophylaxis and treatment of immune of inflammatory disorders, or disorders involving the inappropriate growth or migration of cells.

[0001] This invention relates to a series of squaric acid derivatives,to compositions containing them, to processes for their preparation, andto their use in medicine.

[0002] Over the last few years it has become increasingly clear that thephysical interaction of inflammatory leukocytes with each other andother cells of the body plays an important role in regulating immune andinflammatory responses [Springer, T A. Nature, X, 425, (1990); Springer,T. A. Cell 76, 301, (1994)]. Many of these interactions are mediated byspecific cell surface molecules collectively referred to as celladhesion molecules.

[0003] The adhesion molecules have been sub-divided into differentgroups on the basis of their structure. One family of adhesion moleculeswhich is believed to play a particularly important role in regulatingimmune and inflammatory responses is the integrin family. This family ofcell surface glycoproteins has a typical non-covalently linkedheterodimer structure. At least 14 different integrin alpha chains and 8different integrin beta chains have been identified [Sonnenberg, ACurrent Topics in Microbiology and Immunology, 184, 7, (1993)]. Themembers of the family are typically named according to their heterodimercomposition although trivial nomenclature is widespread in this field.Thus the integrin termed α4β1 consists of the integrin alpha 4 chainassociated with the integrin beta 1 chain, but is also widely referredto as Very Late Antigen 4 or VLA4.

[0004] Some integrin chains are capable of pairing with more than onepartner. For example, the α_(v) chain has been reported to pair with thebeta 1 chain, the beta 3 chain, the beta 5 chain, the beta 6 chain andthe beta 8 chain to give molecules which bind to different sets ofligands and which are referred to respectively as the integrinsα_(v)β₁,α_(v)β₃, α_(v)β₅, α_(v)β₆, and α_(v)β₈. Not all of the potentialpairings of integrin alpha and beta chains have yet been observed innature and the integrin family has been subdivided into a number ofsubgroups based on the pairings that have been recognised [Sonnenberg,A. ibid].

[0005] The importance of integrin function in normal physiologicalresponses is highlighted by two human deficiency diseases in whichintegrin function is defective. Thus in the disease termed LeukocyteAdhesion Deficiency (LAD) there is a defect in one of the families ofintegrins expressed [on leukocytes. Patients suffereing from thisdisease have a reduced ability to recruit leukocytes to inflammatorysites and suffer recurrent infections which in extreme cases may befatal. In the case of patients suffering from the disease termedGlanzman's thrombasthenia,(a defect in a member of the beta 3 integerfamly) there is a defect in blood clotting.

[0006] The potential to modify integrin function in such a way as tobeneficially modulate cell adhesion has been extensively investigated inanimal models using specific monoclonal antibodies and peptides thatblock various functions of these molecules [e.g. Issekutz, T. B. J.Immunol. 3394, (1992); Li, Z. et al. Am. J. Physiol. 263, L723, (1992);Mitjans et al J. Cell Sci. 108, 2825 (1995), Brooks P.C. et al J. Clin.Invest. 96, 1815 (1995), Binns, R. M. et al J. Immunol. 157, 4094,(1996), Hammes, H-P, et al Nature Medicine 2, 529 (1996), Srivata, S. etal Cardiovascular Res. 3, 408 (1997)]. A number of monoclonal antibodieswhich block integrin function are currently being investigated for theirtherapeutic potential in human disease.

[0007] Inhibition of integrin-mediated cell interaction can be expectedto be beneficial in a number of disease states, and in addition to themonoclonal antibodies and peptides just mentioned there has been greatinterest in selective low molecular weight inhibitors of integrinfunction. Thus, for example selective α₄ integrin inhibitors have beendescribed in International patent Specifications Nos. WO96122966,WO97/03094, WO 98/04247, WO98/04913, WO98/53814, WO98/53817, WO98/53818,WO98/54207, WO98/58902, WO99/06390, WO99/06431-06437, WO99/10312,WO99/10313, WO99/67230, WO 99/26922, WO99/60015, WO99/26921, WO9936393,WO99/52898 and WO99/64395. Numerous selective α_(v) integrin inhibitorshave also been described, for example in International PatentSpecifications Nos. WO97/08145, WO97/23480, WO97/36858, WO97/36859,WO97/36861, WO97/36862, WO97/44333, WO97/47618, WO98/31359, WO98/25892,WO98/18460, WO99/44994, WO99/30709, WO99/31061, WO 99126945, WO99152896,WO99/52879, WO99/32457, WO99/31099, WO00/07544, WO00/00486, WO00/06169,WO0/17197 and WO00/01383.

[0008] While it is clearly possible to obtain selective integrininhibitors, their usefulnesses in medicine may be limited due to poorpharmacokinetic properties. Thus, for example, in our hands, integrininhibitors falling within the general structural types featured in theabove-mentioned patent specifications are not particularly attractivefor development as medicines since they can be cleared rapidly from thebody. In order to overcome this problem we have made use of a squaricacid framework which can be readily adapted to provide potent andselective integrin inhibitors using recognised integrin binding groups(for example as described herein and in the patent specifications listedabove), which advantageously possess good pharmacokinetic properties,especially low plasma clearance.

[0009] Thus according to one aspect of the invention we provide acompound of formula (1)

[0010] wherein

[0011] R¹ is an integrin binding group;

[0012] R² is a hydrogen atom or a C₁₋₆alkyl group;

[0013] L¹ is a covalent bond or a linker atom or group;

[0014] n is zero or the integer 1;

[0015] Alk¹ is an optionally substituted aliphatic chain;

[0016] R³ is a hydrogen atom or an optionally substitutedheteroaliphatic, cycloaliphatic, heterocycloaliphatic,polycycloaliphatic, polyheterocycloaliphatic, aromatic or heteroaromaticgroup:

[0017] and the salts, solvates, hydrates and N-oxides thereof.

[0018] It will be appreciated that compounds of formula (1) may have oneor more chiral centres, and exist as enantiomers or diastereomers. Theinvention is to be understood to extend to all such enantiomers,diastereomers and mixtures thereof, including racemates. Formula (1) andthe formulae hereinafter are intended to represent all individualisomers and mixtures thereof. unless stated or shown otherwise.

[0019] In the compounds according to the invention, integrin-bindinggroups represented by R¹ include for example those which are able tobind α₄- or α_(v)-integrins. Particular examples of such integrinsinclude α₄β₁, α₄β₇ and α_(v)β₃ integrins.

[0020] In general, the term integrin-binding group is used herein inrelation to R¹ to mean any group which when part of the compound offormula (1) is able to interact with an integrin to modulate celladhesion in vivo and achieve a therapeutic response. Typically the R¹group may bind to the integrin in such a way that it modulates theinteraction of the integrin with its ligand. Thus for example the R¹group may inhibit binding of the ligand and decrease cell adhesion. Sucha response enables appropriate R¹ groups to be readily identified usingsmall scale routine in vitro screening assays to determine the degree ofinhibition of integrin-ligand binding in the presence of a compound offormula (1). Examples of such screening assays are widely reported inthe literature, for example in the papers and International patentspecifications described above, and in the Examples hereinafter.

[0021] Thus in general R¹ may be any group which when present in acompound of formula (1) is able to bind to an integrin such that thecompound of formula (1) inhibits the binding of the integrin with itsligand with an IC₅₀ of 10 μM or below, particularly 1 μM or below,especially 500 nM or below, e.g. in the range 0.001-500 nM.

[0022] Particular R¹ groups in compounds of the invention include thoseof formula Ar¹L²Ar²Alk- wherein Ar¹ is an optionally substitutedaromatic or heteroaromatic group, L² is a linker atom or group, Ar² isan optionally substituted phenylene or nitrogen-containing six-memberedheteroarylene group and Alk is a chain:

[0023] where R is a carboxylic acid (—CO₂H) or a derivative or biosterethereof. R¹ groups of this type are particularly useful for binding α₄integrins and compounds of formula (1) incorporating the Ar₁L²Ar²Alk-function can be expected to inhibit (α₄ integrins such as α₄β₁ and/orα₄β₇ at concentrations at which they generally have no or minimalinhibitory action on integrins of other ax subgroups. Such compounds areof use in medicine, for example in the prophylaxis and treatment ofimmune or inflammatory disorders as described hereinafter.

[0024] Optionally substituted aromatic groups represented by Ar₁ whenpresent in the group R¹ include for example optionally substitutedmonocyclic or bicyclic fused ring C₆₋₁₂ aromatic groups, such as phenyl,1- or 2-naphthyl, 1- or 2-tetrahydronaphthyl, indanyl or indenyl groups.

[0025] Optionally substituted heteroaromatic groups represented by thegroup Ar¹ when present in the group R¹ include for example optionallysubstituted C₁₋₉ heteroaromatic groups containing for example one, two,three or four heteroatoms selected from oxygen, sulphur or nitrogenatoms. In general, the heteroaromatic groups may be for examplemonocyclic or bicyclic fused ring, heteroaromatic groups. Monocyclicheteroaromatic groups include for example five- or six-memberedheteroaromatic groups containing one, two, three or four heteroatomsselected from oxygen, sulphur or nitrogen atoms. Bicyclic heteroaromaticgroups include for example eight- to thirteen-membered fused-ringheteroaromatic groups containing one, two or more heteroatoms selectedfrom oxygen, sulphur or nitrogen atoms.

[0026] Particular examples of heteroaromatic groups of these typesinclude pyrrolyl, furyl, thienyl, imidazolyl, N—C₁₋₁₆alkylimidazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl,1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,3,4-thiadiazole, pyridyl,pyrimidinyl, pyridazinyl, pyrazinyl, 1,3,5-triazinyl, 1,2,4-triazinyl,1,2,3-triazinyl, benzofuryl, [2,3-dihydro]benzofuryl,[2,3-dihydro]benzothienyl, benzothienyl, benzotriazolyl, indolyl,isoindolyl, benzimidazolyl, imidazo[1,2-a]pyridyl, benzothiazolyl,benzoxazolyl, benzopyranyl, [3,4-dihydro]benzopyranyl, quinazolinyl,qiunoxalinyl, naphthyridinyl, especially 2,6-naphthyridinyl,pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl, pyrido[4,3-b]-pyridyl,quinolinyl, isoquinolinyl, tetrazolyl, 5,6,7,8-tetrahydroquinolinyl,5,6,7,8-tetrahydroisoquinolinyl, and imidyl, e.g. succinimidyl,phthalimidyl, or naphthalimidyl such as 1,8-naphthalimidyl.

[0027] Each aromatic or heteroaromatic group represented by the groupAr¹ may be optionally substituted on any available carbon or, whenpresent, nitrogen atom. One, two, three or more of the same or differentsubstituents may be present and each substituent may be selected forexample from an atom or group -L³(Alk²)_(t)L⁴(R⁴)_(u) in which L³ andL⁴, which may be the same or different, is each a covalent bond or alinker atom or group, t is zero or the integer 1, u is an integer 1, 2or 3, Alk² is an aliphatic or heteroaliphatic chain and R⁴ is a hydrogenor halogen atom or a group selected from optionally substitutedC₁₋₆alkyl or C₃₋₈ cycloalkyl, —OR⁵ [where R⁵ is a hydrogen atom, anoptionally substitued C₁₋₆alkyl or C₃₋₈ cycloalkyl group], —SR⁵, —NR⁵R⁶[where R⁶ is as just defined for R⁵ and may be the same or different],—NO₂, —CN, —CO₂R⁵, —SO₃H, —SOR⁵, —SO₂R⁵, —SO₃R⁵, —OCO₂R⁵, —CONR⁵R⁶,—OCONR⁵R⁶, —CSNR⁵R⁶—COR⁵, —OCOR⁵, —N(R⁵)COR⁶, —N(R⁵)CSR⁶, —SO₂N(R⁵)(R⁶),—N(R⁵)SO₂RS, N(R⁵)CON(R⁶)(R⁷) [where R⁷ is a hydrogen atom, anoptionally substituted C₁₋₆alkyl or C₃₋₈cycloalkyl group],—N(R⁵)CSN(R⁶)(R⁷) or —N(R⁵)SO₂N(R⁶)(R⁷), provided that when t is zeroand each of L³ and L⁴ is a covalent bond then u is the integer 1 and R⁴is other than a hydrogen atom.

[0028] When L³ and/or L⁴ is present in these substituents as a linkeratom or group it may be any divalent linking atom or group. Particularexamples include —O— or —S— atoms or —C(O)—, —C(O)O—, —OC(O)—, —C(S)—,—S(O)—, —S(O)₂—, —N(R⁸)-[where R⁸ is a hydrogen atom or an optionallysubstituted C₁₋₆alkyl group], —N(R⁸)O—, —N(R⁸)N— —CON(R⁸)—,—OC(O)N(R⁸)—, —CSN(R⁸)—, —N(R⁸)CO—, —N(R⁸)C(O)O—, —N(R⁸)CS—,—S(O)₂N(R⁸)—, —N(R⁸)S(O)₂—, —N(R⁸)CON(R⁸)—, —N(R⁸)CSN(R⁸)—, or—N(R⁸)SO₂N(R⁸)— groups. Where the linker group contains two R⁸substituents, these may be the same or different.

[0029] When R⁴, R⁵, R⁶, R⁷ and/or R⁸ is present as a C₁₋₆alkyl group itmay be a straight or branched C₁₋₆alkyl group, e.g. a C₁₋₃alkyl groupsuch as a methyl or ethyl group. C₃₋₈cycloalkyl groups represented byR⁴, R⁵, R⁶, R⁷ and/or R⁸ include C₃₋₆cycloalkyl groups e.g. cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl groups. Optional substituentswhich may be present on such alkyl and cycloalkyl groups include forexample one, two or three substituents which may be the same ordifferent selected from halogen atoms, for example fluorine, chlorine,bromine or iodine atoms, or hydroxy or C₁₋₆alkoxy e.g. methoxy or ethoxygroups.

[0030] When the groups R⁵ and R⁶ or R⁶ and R⁷ are both C₁₋₆alkyl groupsthese groups may be joined, together with the N atom to which they areattached, to form a heterocyclic ring. Such heterocyclic rings may beoptionally interrupted by a further heteroatom selected from —O—, —S— or—N(R⁵)—. Particular examples of such heterocyclic rings includepiperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl, imidazolidinyland piperazinyl rings.

[0031] When Alk² is present as an aliphatic or heteroaliphatic chain itmay be for example any, divalent chain corresponding to thebelow-mentioned aliphatic or heteroaliphatic group described for Alk¹ orR³ respectively.

[0032] Halogen atoms represented by R⁴ in the optional Ar¹ substituentsinclude fluorine, chlorine, bromine, or iodine atoms.

[0033] Examples of the substituents represented by-L³(Alk²)_(t)L⁴(R⁴)_(u) when present in Ar¹ groups in compounds of theinvention include atoms or groups -L³Alk²L⁴R⁴, -L³Alk²R⁴, -L³R⁴, —R⁴ and-Alk²R⁴ wherein L³, Alk², L⁴ and R⁴ are as defined above. Particularexamples of such substituents include -L³CH₂L⁴R⁴, -L³CH(CH₃)L⁴R⁴,-L³CH(CH₂)₂L⁴R⁴, -L³CH₂R⁴, -L³CH(CH₃)R⁴, -L³(CH₂)₂R⁴, —CH₂R⁴,—CH(CH₃)R⁴, —(CH₂)₂R⁴ and —R⁴ groups.

[0034] Thus Ar¹ in compounds of the invention may be optionallysubstituted for example by one, two, three or more halogen atoms, e.g.fluorine, chlorine, bromine or iodine atoms, and/or C₁₋₆alkyl, e.g.methyl, ethyl, n-propyl, i-propyl, n-butyl or t-butyl, C₃₋₈cycloalkyl,e.g. cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl,C₁₋₆hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl or —C(OH)(CF₃)₂,carboxyC₁₋₆alkyl, e.g. carboxyethyl, C₁₋₆alkylthio e.g. methylthio orethylthio, carboxyC₁₋₆alkylthio, e.g. carboxymethylthio,2-carboxyethylthio or 3-carboxy-propylthio, C₁₋₆alkoxy, e.g. methoxy orethoxy, hydroxyC₁₋₆alkoxy, e.g. 2-hydroxyethoxy, haloC₁₋₆alkyl, e.g.—CF₃, —CHF₂, CH₂F, haloC₁₋₆alkoxy, e.g. —OCF₃, —OCHF₂, —OCH₂F,C₁₋₆alkylamino, e.g. methylamino or ethylamino, amino (—NH₂),aminoC₁₋₆alkyl, e.g. aminomethyl or aminoethyl, C₁₋₆dialkylamino, e.g.dimethylamino or diethylamino, C₁₋₆alkylaminoC₁₋₆alkyl, e.g.ethylaminoethyl, C₁₋₆ dialkylaminoC₁₋₆alkyl, e.g. diethylaminoethyl,aminoC₁₋₆alkoxy, e.g. aminoethoxy, C₁₋₆alkylaminoC₁₋₆alkoxy, e.g.methylaminoethoxy, C₁₋₆dialkylaminoC₁₋₆alkoxy, e.g. dimethylaminoethoxy,diethylaminoethoxy, isopropylaminoethoxy, or dimethylaminopropoxy,nitro, cyano, amidino, hydroxyl (—OH), formyl [HC(O)-], carboxyl(—CO₂H), —CO₂Alk³ (where Alk³ is as defined below for Alk⁷], C₁₋₆alkanoyl e.g. acetyl, thiol (—SH), thioC₁₋₆alkyl, e.g. thiomethyl orthioethyl, sulphonyl (—SO₃H), —SO₃Alk³, C₁₋₆alkylsulphinyl, e.g.methylsulphinyl, C₁₋₆alkylsulphonyl, e.g. methylsulphonyl,aminosulphonyl (—SO₂NH₂), C₁₋₆ alkylaminosulphonyl, e.g.methylaminosulphonyl or ethylaminosulphonyl, C₁₋₆dialkylaminosulphonyl,e.g. dimethylaminosulphonyl or diethylaminosulphonyl,phenylaminosulphonyl, carboxamido (—CONH₂), C₁₋₆alkyl aminocarbonyl,e.g. methylaminocarbonyl or ethylaminocarbonyl,C₁₋₆dialkylaminocarbonyl, e.g. dimethylaminocarbonyl ordiethylaminocarbonyl, aminoC₁₋₆alkylaminocarbonyl, e.g.aminoethylaminocarbonyl, C₁₋₆dialkylaminoC₁₋₆alkylaminocarbonyl, e.g.diethylaminoethylaminocarbonyl, aminocarbonylamino,C₁₋₆alkylaminocarbonylamino, e.g. methylaminocarbonylamino orethylaminocarbonylamino, C₁₋₆dialkylaminocarbonylamino, e.g.dimethylaminocarbonylamino or diethylaminocarbonylamino,C₁₋₆alkylaminocabonylC₁₋₆ alkylamino, e.g.methylaminocarbonylmethylamino aminothiocarbonylamino,C₁₋₆alkylaminothiocarbonylamino, e.g. methylaminothiocarbonylamino orathylaminothiocarbonylamino, C₁₋₆dialkylaminothiocarbonylamino, e.g.dimethylaminothiocarbonylamino or diethylaminothiocarbonylamino,C₁₋₆alkylaminothiocarbonylC₁₋₆alkylamino, e.g.ethylaminothiocarbonylmethylamino, C₁₋₆alkylsulphonytamino, e.g.methylsulphonylamino or ethylsulphonylemino, C₁₋₆dialkylsulphonylamino,e.g. dimethylsulphonylamino or diethyl-sulphonylamino,aminosulphonylamino (—NHSO₂NH₂), C₁₋₆alkylamino-sulphonylamino, e.g.methylaminosulphonylamino or ethylaminosulphonylamino,C₁₋₆dialkylaminosulphonylamino, e.g. dimethylaminosulphonylamino ordiethylaminosulphonylamino, C₁₋₆alkanoylamino, e.g. acetylamino,aminoC₁₋₆alkanoylamino e.g. aminoocetylamino,C₁₋₆dialkylaminoC₁₋₆alkanoylamino, e.g. dimethylaminoacetylamino,C₁₋₆alkanoylaminoC₁₋₆alkyl, e.g. acetylaminomethyl,C₁₋₆alkanoylaminoC₁₋₆alkylamino, e.g. acetamidoethylamino,C₁₋₆alkoxycarbonylamino, e.g: methoxycarbonylamino, ethoxycarbonylaminoor t-butoxycarbonylamino groups.

[0035] L² when present as part of the group R¹ in compounds of theinvention may be a linker atom or group L^(2a) or a linker-Alk^(a)(L^(2a))_(y)-, where Alk^(a) is an optionally substitutedaliphatic or heteroaliphatic chain as previously defined for Alk², andL^(2a) is a linker atom or group as described above for L³ and L⁴ and yis zero or the integer 1.

[0036] Optionally substituted nitrogen-containing six-memberedheteroarylene groups represented by Ar² when present as part of thegroup R¹ include optionally substituted pyridiyl, pyrimidindiyl,pyridazindiyl, pyrazindiyl and triazindiyl groups. Each group may beattached to the remainder of the molecule through any available ringcarbon atoms.

[0037] The phenylene and nitrogen-containing heteroarylene groupsrepresented by Ar² may be optionally substituted by one or twosubstituents selected from the atoms or groups -L³(Alk²)_(t)L⁴(R⁴)_(u)described herein. Where two of these atoms or groups are present theymay be the same or different.

[0038] When the group R is present in R¹ in compounds of the inventionas a derivative of a carboxylic acid it may be for example a carboxylicacid ester or amide. Particular esters and amides include —CO₂Alk⁷ and—CONR⁵R⁶ groups as defined herein. When R is a biostere of a carboxylicacid it may be for example a tetrazole or other acid such as phosphonicacid, phosphinic acid, sulphonic acid, sulphinic acid or boronic acid oran acylsulphonamide group.

[0039] When the group R² is present in compounds of the invention as aC₁₋₆alkyl group it may be for example a straight or branched C₁₋₆alkylgroup, e.g. a C₁₋₃alkyl group such as a methyl or ethyl group.

[0040] The linker atom or group represented by L¹ in compounds offormula (1) may be any linker atom or group as described above for thelinker atom or group L³.

[0041] When the group Alk¹ is present in compounds of formula (1) as anoptionally substituted aliphatic chain it may be an optionallysubstituted C₁₋₁₀ aliphatic chain. Particular examples includeoptionally substituted straight or branched chain C₁₋₆ alkylene, C₂₋₆alkenylene, or C₂₋₆ alkynylene chains.

[0042] Particular examples of aliphatic chains represented by Alk¹include optionally substituted —CH₂—, —(CH₂)₂—, —CH(CH₃)CH₂—,—(CH₂)₂CH₂—, —(CH₂)₃CH₂—, —CH(CH₃)(CH₂)₂—, —CH₂CH(CH₃)CH₂—,—C(CH₃)₂CH₂—, —CH₂C(CH₃)₂CH₂—, —(CH₂)₂C(CH₃)₂CH₂—, —(CH₂)₄CH₂—,—(CH₂)₅CH₂—, —CHCH—, —CHCHCH₂—, —CH₂CHCH—, —CHCHCH₂CH₂—, —CH₂CHCHCH₂—,—(CH₂)₂CHCH—, —CC—, —CCCH₂—, —CH₂CC—, —CCCH₂CH₂—, —CH₂CCCH₂— or—(CH₂)₂CCH— groups.

[0043] Heteroaliphatic groups represented by the group R³ in thecompounds of formula (1) include the aliphatic chains just described forAlk¹ but with each containing a terminal hydrogen atom and additionallycontaining one, two, three or four heteroatoms or heteroatom-containinggroups. Particular heteroatoms or groups include atoms or groups L⁵where L⁵ is as defined above for L³ when L³ is a linker atom or group.Each L⁵ atom or group may interrupt the aliphatic group, or may bepositioned at its terminal carbon atom to connect the group to anadjoining atom or group. Particular examples include optionallysubstituted -L⁵CH₃, —CH₂L⁵CH₃, -L⁵CH₂CH₃, —CH₂L⁵CH₂CH₃, —(CH₂)₂L⁵CH₃,—(CH₂)₃L⁵CH₃, -L⁵(CH₂)₃, and —(CH₂)₂L⁵CH₂CH₃ groups.

[0044] The optional substituents which may be present on aliphatic orheteroaliphatic chains represented by Alk¹ and R³ respectively includeone, two, three or more substituents where each substituent may be thesame or different and is selected from halogen atoms, e.g. fluorine,chlorine, bromine or iodine atoms, or —OH, —CO₂H, —CO₂R⁹, where R⁹ is anoptionally substituted straight or branched C₁₋₆alkyl group as definedabove for R⁴, —CONHR⁹, —CON(R^(a))₂, —COCH₃, C₁₋₆alkoxy, e.g. methoxy orethoxy, thiol, —S(O)R⁹, —S(O)₂R⁹, C₁₋₆alkylthio e.g. methylthio orethylthio, amino or substituted amino groups. Substituted amino groupsinclude —NHR⁹ and —N(R⁹)₂ groups. Where two R⁹ groups are present in anyof the above substituents these may be the same or different.

[0045] Optionally substituted cycloaliphatic groups represented by thegroup R³ in compounds of the invention include optionally substitutedC₃₋₁₀ cycloaliphatic groups. Particular examples include optionallysubstituted C₃₋₁₀ cycloalkyl, e.g. C₃₋₇ cycloalkyl or C₃₋₁₀cycloalkenyl, e.g C₃₋₇ cycloalkenyl groups.

[0046] Optionally substituted heterocycloaliphatic groups represented bythe group R³ include optionally substituted C₃₋₁₀heterocycloaliphaticgroups. Particular examples include optionally substitutedC₃₋₁₀heterocycloalkyl, e.g. C₃₋₇ heterocycloalkyl, orC₃₋₁₀heterocycloalkenyl, e.g. C₃₋₇ hetercycloalkenyl groups, each ofsaid groups containing one, two, three or four heteroatoms orheteroatom-containing groups L⁵ as defined above.

[0047] Optionally substituted polycycloaliphatic groups represented bythe group R³ include optionally substitued C₇₋₁₀ bi- or tricycloalkyl orC₇₋₁₀bi- or tricycloalkenyl groups. Optionally substitutedpolyheterocycloaliphatic groups represented by the group R³ include theoptionally substituted polycycloalkyl groups just described, but witheach group additionally containing one, two, three or four L⁵ atoms orgroups.

[0048] Particular examples of cycloaliphatic, polycycloaliphatic,heterocycloaliphatic and polyheterocycloaliphatic groups represented bythe group R³ include optionally substituted cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, 2-cyclobuten-1-yl,2-cyclopenten-1-yl, 3-cyclopenten-1-yl, adamantyl, norbornyl,norbornenyl, tetrahydrofuranyl, pyrroline, e.g. 2- or 3-pyrrolinyl,pyrrolidinyl, pyrrolidinone, oxazolidinyl, oxazolidinone, dioxolanyl,e.g. 1,3-dioxolanyl, imidazolinyl, e.g. 2-imidazolinyl, imidazolidinyl,pyrazolinyl, e.g. 2-pyrazolinyl, pyrazolidinyl, pyranyl, e.g. 2- or4-pyranyl, piperidinyl, piperidinone, 1,4-dioxanyl, morpholinyl,morpholinone, 1,4-dithianyl, thiomorpholinyl, piperazinyl,1,3,5-trithianyl, oxazinyl, e.g. 2H-1,3-, 6H-1,3-, 6H-1,2-, 2H-1,2- or4H-1,4-oxazinyl, 1,2,5-oxathiazinyl, isoxazinyl, e.g. o- orp-isoxazinyl, oxathiazinyl, e.g. 1,2,5 or 1,2,6-oxathiazinyl, or1,3,5,-oxadiazinyl groups.

[0049] The optional substituents which may be present on thecycloaliphatic, polycycloaliphatic, heterocycloaliphatic orpolyheterocycloaliphatic groups represented by the group R³ include one,two, three or more substituents each selected from halogen atoms, e.g.fluorine, chlorine, bromine or iodine atoms, or C₁₋₆alkyl, e.g. methylor ethyl, haloC₁₋₆alkyl, e.g. halomethyl or haloethyl such asdifluoromethyl or trifluoromethyl, optionally substituted by hydroxyl,e.g. —C(OH)(CF₃)₂, C₁₋₆alkoxy, e.g. methoxy or ethoxy, haloC₁₋₆alkoxy,e.g. halomethoxy or haloethoxy such as difluoromethoxy ortrifluoromethoxy, thiol, C₁₋₆alkylthio e.g. methylthio or ethylthio, or-(Alk⁴)_(v)R¹⁰ groups in which Alk⁴ is a straight or branchedC₁₋₃alkylene chain, v is zero or an integer 1 and R¹⁰ is a —OH, —SH,—N(R¹¹)₂, (in which R¹¹ is an atom or group as defined herein for R⁸)—CN, —CO₂R¹¹, —NO₂, —CON(R¹¹)₂, —CSN(R¹¹)₂, —COR¹¹, —CSN(R¹¹)₂,—N(R¹¹)COR¹ 1, —N(R¹¹)CSR¹¹, —SO₂N(R¹¹)₂, —N(R 1)SO₂R¹¹,—N(R¹¹)CON(R¹¹)₂, —N(R¹¹)CSN(R¹¹), N(R¹¹)SO₂N(R¹¹)₂ or optionallysubstituted phenyl group. Where two R¹¹ atoms or groups are present inthese substituents these may be the same or different. Optionallysubstituted phenyl groups include phenyl substituted by one, two orthree of the R¹³ groups described below

[0050] Additionally, when the group R³ is a heterocycloaliphatic groupcontaining one or more nitrogen atoms each nitrogen atom may beoptionally substituted by a group -(L⁶)_(p)(Alk⁵)_(q)R¹² in which L⁶ is—C(O)—, —C(O)O—, —C(S)—, —S(O)₂—, —CON(R¹¹)—, —CSN(R¹¹)— or SO₂N(R¹¹)—;p is zero or an integer 1; Alk⁵ is an optionally substituted aliphaticor heteroaliphatic chain; q is zero or an integer 1; and R¹² is ahydrogen atom or an optionally substituted cycloaliphatic,heterocycloaliphatic, polycycloaliphatic, polyheterocycloaliphatic,aromatic or heteroaromatic group.

[0051] Optionally substituted aliphatic or heteroaliphatic chainsrepresented by Alk⁵ include those optionally substituted chainsdescribed above for Alk¹ and R³ respectively.

[0052] Cycloaliphatic, heterocycloaliphatic, polycycloaliphatic orpolyheterocycloaliphatic groups represented by R¹² include those groupsjust described for the group R³. Optional substituents which may bepresent on these groups include those described above in relation toAlk¹ and R³ aliphatic and heteroaliphatic chains.

[0053] When the group R³ is an optionally substituted aromatic orheteroaromatic group it may be for example an aromatic or heteroaromaticgroup as described herein for the group Ar¹.

[0054] Optional substituents which may be present on the aromatic orheteroaromatic, groups represented by the group R³ include one, two,three or more substituents, each selected from an atom or group R¹³ inwhich R¹³ is —R^(13a) or -Alk⁶(R^(13a))_(m), where R^(13a) is a halogenatom, or an amino (—NH₂), substituted amino, nitro, cyano, amidino,hydroxyl (—OH), substituted hydroxyl, formyl, carboxyl (—CO₂H),esterified carboxyl, thiol (—SH), substituted thiol, —COR¹⁴ [where R¹⁴is an -Alk⁶(R^(13a))_(m), cycloaliphatic, heterocycloaliphatic, aryl orheteroaryl group], —CSR¹⁴, —SO₃H, —SOR¹⁴, —SO₂R¹⁴, —SO₃R¹⁴, —SO₂NH₂,—SO₂NHR¹⁴ SO₂N(R¹⁴)₂, —CONH₂, —CSNH₂, —CONHR¹⁴, —CSNHR¹⁴, —CON[R¹⁴]2,—CSN(R¹⁴)₂, —N(R¹¹)SO₂R¹⁴, —N(SO₂R¹⁴)₂, —NH(R¹¹)SO₂NH₂, —N(R¹¹)SO₂NHR¹⁴,—N(R¹¹)SO₂N(R¹⁴)₂, —N(R¹¹)COR¹⁴, —N(R¹¹)CONH₂, —N(R¹¹)CONHR¹⁴,—N(R¹¹)CON(R¹⁴), —N(R¹¹)CSNH₂, —N(R¹¹)CSNHR¹⁴, —N(R¹¹)CSN(R¹⁴)₂,—N(R¹¹)CSR¹⁴, —N(R¹¹)C(O)OR¹⁴, —SO₂NHet¹ [where —NHet¹ is an optionallysubstituted C₅₋₇cyclicamino group optionally containing one or moreother —O— or —S— atoms or —N(R¹¹)—, —C(O)—, —C(S)—, S(O) or —S(O)₂groups], —CONHet¹, —CSNHet¹, —N(R¹¹)SO₂NHet¹, —N(R¹¹)CONHet¹,—N(R¹¹)CSNHet¹. —SO₂N(R¹¹)Het² [where Het² is an optionally substitutedmonocyclic C₅₋₇carbocyclic group optionally containing one or more —O—or —S— atoms or —N(R¹¹)—, —C(O)— or —C(S)— groups], -Het²,—CON(R¹¹)Het², —CSN(R¹¹)Het², —N(R¹¹)CON(R¹¹)Het², —N(R¹¹)CSN(R¹¹)Het²,cycloaliphatic, heterocycloaliphatic, aryl or heteroaryl group; Alk⁶ isa straight or branched C₁₋₆alkylene, C₂₋₆alkenylene or C₂₋₆alkynylenechain, optionally interrupted by one, two or three —O— or —S— atoms orS(O)_(n) [where n is an integer 1 or 2] or —N(R¹⁵)— groups [where R¹⁵ isa hydrogen atom or C₁₋₆alkyl, e.g. methyl or ethyl group]; and m is zeroor an integer 1, 2 or 3. It will be appreciated that when two R¹¹ or R¹⁴groups are present in one of the above substituents, the R¹¹ or R¹⁴groups may be the same or different.

[0055] When in the group -Alk⁶(R^(13a))_(m) m is an integer 1, 2 or 3,it is to be understood that the substituent or substituents R^(13a) maybe present on any suitable carbon atom in -Alk⁶. Where more than oneR^(13a) substituent is present these may be the same or different andmay be present on the same or different atom in -Alk⁶. Clearly, when mis zero and no substituent R^(13a) is present the alkylene, alkenyleneor alkynylene chain represented by Alk⁶ becomes an alkyl, alkenyl oralkynyl group.

[0056] When R^(13a) is a substituted amino group it may be for example agroup —NHR¹⁴ [where R¹⁴ is as defined above] or a group —N(R¹⁴2 whereineach R¹⁴ group is the same or different.

[0057] When R^(13a) is a halogen atom it may be for example a fluorine,chlorine, bromine, or iodine atom.

[0058] When R^(13a) is a substituted hydroxyl or substituted thiol groupit may be for example a group —OR¹⁴ or a —SR¹⁴ or —SC(═NH)NH₂ grouprespectively. Esterified carboxyl groups represented by the groupR^(13a) include groups of formula —CO₂Alk⁷ wherein Alk⁷ is a straight orbranched, optionally substituted C₁₋₈alkyl group such as a methyl,ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl or t-butyl group; aC₆₋₁₂arylC₁₋₈alkyl group such as an optionally substituted benzyl,phenylethyl, phenylpropyl, 1-naphthylmethyl or 2-naphthylmethyl group; aCeC₁₋₂aryl group such as an optionally substituted phenyl, 1-naphthyl or2-naphthyl group; a C₆₋₁₂aryloxyC₁₋₈alkyl group such as an optionallysubstituted phenyloxymethyl, phenyloxyethyl, 1-naphthyloxymethyl, or2-naphthyloxymethyl group; an optionally substitutedC₁₋₈alkanoyloxyC₁₋₈alkyl group, such as a pivaloyloxymethyl,propionyloxyethyl or propionyloxypropyl group; or aC₆₋₁₂aroyloxyC₁₋₁₈alkyl group such as an optionally substitutedbenzoyloxyethyl or benzoyloxypropyl group. Optional substituents presenton the Alk⁷ group include R^(13a) substituents described above.

[0059] When Alk⁶ is present in or as a substituent it may be for examplea methylene, ethylene, n-propylene, i-propylene, n-butylene, i-butylene,s-butylene, t-butylene, ethenylene, 2-propenylene, 2-butenylene,3-butenylene, ethynylene, 2-propynylene, 2-butynylene or 3-butynylenechain, optionally interrupted by one, two, or three —O— or —S—, atoms or—S(O)—, —S(O)₂— or —N(R⁹)— groups. Cycloaliphatic orheterocycloaliphatic groups represented by the groups R^(13a) or R¹⁴include those optionally substituted C₃₋₁₀cycloaliphatic or C₃₋₁₀heterocycloaliphatic groups described above for R³.

[0060] Aryl or heteroaryl groups represented by the groups R^(13a) orR¹⁴ include mono- or bicyclic optionally substituted C₆₋₁₂ aromatic orC₁₋₉ heteroaromatic groups as described above for the group Ar¹. Thearomatic and heteroaromatic groups may be attached to the remainder ofthe compound of formula (1) by any carbon or hetero e.g. nitrogen atomas appropriate.

[0061] When —NHet¹ or -Het² forms part of a substituent R¹³ each may befor example an optionally substituted pyrrolidinyl, pyrazolidinyl,piperazinyl, morpholinyl, thiomorpholinyl, piperidinyl or thiazolidinylgroup. Additionally Het² may represent for example, an optionallysubstituted cyclopentyl or cyclohexyl group. Optional substituents whichmay be present on —NHet¹ or -Het² include those R⁷ substituentsdescribed above.

[0062] Particularly useful atoms or groups represented by R¹³ includefluorine, chlorine, bromine or iodine atoms, or C₁₋₆alkyl, e.g. methyl,ethyl, n-propyl, i-propyl, n-butyl or t-butyl, optionally substitutedphenyl, pyridyl, pyrimidinyl, pyrrolyl, furyl, thiazolyl, thienyl,morpholinyl, thiomorpholinyl, piperazinyl, e.g.t-butyloxycarbonylpiperazinyl, pyrrolidinyl, dioxolanyl, dioxanyl,oxazolidinyl, thiazolidinyl, imidazolidinyl or piperidinyl,C₁₋₆hydroxyalkyl, e.g. hydroxymethyl or hydroxyethyl, carboxyC₁₋₆alkyl,e.g. carboxyethyl, C₁₋₆alkylthio e.g. methylthio or ethylthio,carboxyC₁₋₆alkylthio, e.g. carboxymethylthio, 2-carboxyethylthio or3-carboxypropylthio, C₁₋₆alkoxy, e.g. methoxy or ethoxy,hydroxyC₁₋₆alkoxy, e.g. 2-hydroxyethoxy, optionally substituted phenoxy,pyridyloxy, thiazolyoxy, phenylthio or pyridylthio, C₄₋₇cycloalkyl, e.g.cyclobutyl, cyclopentyl, C₅₋₇cycloalkoxy, e.g. cyclopentyloxy,haloC₁₋₁₆alkyl, e.g. trifluoromethyl, haloC₁₋₆alkoxy, e.g.trifluoromethoxy, C₁₋₆alkylamino, e.g. methylamino, ethylamino orpropylamino, C₆₋₁₂arylC₁₋₆alkylamino, e.g. benzylamino,4-fluorobenzylamino or 4-hydroxyphenylethylamino, amino (—NH₂),aminoC₁₋₆alkyl, e.g. aminomethyl or aminoethyl, C₁₋₆dialkylamino, e.g.dimethylamino or diethylamino, aminoC₁₋₆alkylamino, e.g. aminoethylaminoor aminopropylamino, optionally substituted Het¹NC₁₋₆alkylamino, e.g.3-morpholinopropylamino, C₁₋₆alkylaminoC₁₋₆alkyl, e.g. ethylaminoethyl,C₁₋₆dialkylaminoC₁₋₆alkyl, e.g. diethylaminoethyl, aminoC₁₋₆alkoxy, e.g.aminoethoxy, C₁₋₆alkylaminoC₁₋₆alkoxy, e.g. methylaminoethoxy,C₁₋₆dialkylaminoC₁₋₆alkoxy, e.g. dimethylaminoethoxy,diethylaminoethoxy, diisopropylaminoethoxy, or dimethylaminopropoxy,hydroxyC₁₋₆alkylamino, e.g. 2-hydroxyethylamino, 3-hydroxypropylamino or3-hydroxybutylamino, imido, such as phthalimido or naphthalimido, e.g.1,8-naphthalimido, nitro, cyano, amidino, hydroxyl (—OH), formyl[HC(O)—], carboxyl (—CO₂H), —CO₂Alk⁷ [where Alk⁷ is as defined above],C₁₋₆ alkanoyl e.g. acetyl, propyryl or butyryl, optionally substitutedbenzoyl, thiol (—SH), thioC₁₋₆alkyl, e.g. thiomethyl or thioethyl,—SC(═NH)NH₂, sulphonyl (—SO₃H), —SO₃Alk⁷, C₁₋₆alkylsulphinyl, e.g.methylsulphinyl, ethylsulphinyl or propylsulphinyl, C₁₋₆alkylsulphonyl,e.g. methylsulphonyl, ethylsulphonyl or propylsulphonyl, aminosulphonyl(—SO₂NH₂), C₁₋₆alkylaminosulphonyl, e g methylaminosulphonyl,ethylaminosulphonyl or propylaminosulphonyl C₁₋₆dialkylaminosulphonyl,e.g. dimethylaminosulphonyl or diethylaminosulphonyl,phenylaminosulphonyl, carboxamido (—CONH₂), C₁₋₆alkylaminocarbonyl, e.g.methylaminocarbonyl, ethylaminocarbonyl or propylaminocarbonyl,C₁₋₆dialkylaminocarbonyl, e.g. dimethylaminocarbonyl ordiethylaminocarbonyl, aminoC₁₋₆alkylaminocarbonyl, e.g.aminoethylaminocarbonyl, C₁₋₆alkylaminoC₁₋₆alkylaminocarbonyl, e.g.methylaminoethylaminocarbonyl, C₁₋₆dialkylaminoC₁₋₆alkylaminocarbonyl,e.g. diethylaminoethylaminocarbonyl, aminocarbonylamino,C₁₋₆alkylaminocarbonylamino, e.g. methylaminocarbonylamino orethylaminocarbonylamino, C₁₋₆dialkylaminocarbonylamino, e.g.dimethylaminocarbonylamino or diethylaminocarbonylamino,C₁₋₆alkylaminocabonylC₁₋₆alkylamino, e.g.methylaminocarbonylmethylamino, aminothiocarbonylamino,C₁₋₆alkylaminothiocarbonylamino, e.g. methylaminothiocarbonylamino orethylaminothiocarbonylamino, C₁₋₆dialkylaminothiocarbonylamino, e.g.dimethylaminothiocarbonylamino or diethylaminothiocarbonylamino,C₁₋₆alkylaminothiocarbonylC₁₋₆alkylamino, e.g.ethylaminothiocarbonylmethylamino, —CONHC(═NH)NH₂,C₁₋₆alkylsulphonylamino, e.g. methylsulphonylamino orethylsulphonylamino, haloC₁₋₆alkylsulphonylamino, e.g.trifluoromethylsulphonylamino, C₁₋₆dialkylsulphonylamino, e.g.dimethylsulphonylamino or diethylsulphonylamino, optionally substitutedphenylsulphonylamino, aminosulphonylamino (—NHSO₂NH₂),C₁₋₆alkylaminosulphonylamino, e.g. methylaminosulphonylamino orethylaminosulphonylamino, C₁₋₆dialkylaminosulphonylamino, e.g.dimethylaminosulphonylamino or diethylaminosulphonylamino, optionallysubstituted morpholinesulphonylamino ormorpholinesulphonylC₁₋₆alkylamino, optionally substitutedphenylaminosulphonylamino, C₁₋₆alkanoylamino, e.g. acetylamino,aminoC₁₋₆alkanoylamino e.g. aminoacetylamino,C₁₋₆dialkylaminoC₁₋₆alkanoylamino, e.g. dimethylaminoacetylamino,C₁₋₆alkanoylaminoC₁₋₆alkyl, e.g. acetylaminomethyl,C₁₋₆alkanoylaminoC₁₋₆alkylamino, e.g. acetamidoethylamino,C₁₋₆alkoxycarbonylamino, e.g. methoxycarbonylamino, ethoxycarbonylaminoor t-butoxycarbonylamino or optionally substituted benzyloxy,pyridylmethoxy, thiazolylmethoxy, benzyloxycarbonylamino,benzyloxycarbonylaminoC₁₋₆alkyl e.g. benzyloxycarbonylaminoethyl,thiobenzyl, pyridylmethylthio or thiazolylmethylthio groups.

[0063] Where desired, two R¹³ substituents may be linked together toform a cyclic group such as a cyclic ether, e.g. a C₁₋₆alkylenedioxygroup such as methylenedioxy or ethylenedioxy.

[0064] It will be appreciated that where two or more R¹³ substituentsare present, these need not necessarily be the same atoms and/or groups.In general, the substituent(s) may be present at any available ringposition in the aromatic or heteroaromatic group represented by R³.

[0065] The presence of certain substituents in the compounds of formula(1) may enable salts of the compounds to be formed. Suitable saltsinclude pharmaceutically acceptable salts, for example acid additionsalts derived from inorganic or organic acids, and salts derived frominorganic and organic bases.

[0066] Acid addition salts include hydrochlorides, hydrobromides,hydroiodides, alkylsulphonates, e.g. methanesulphonates,ethanesulphonates, or isothionates, arylsulphonates, e.g.p-toluenesulphonates, besylates or napsylates, phosphates, sulphates,hydrogen sulphates, acetates, trifluoroacetates, propionates, citrates,maleates, fumarates, malonates, succinates, lactates, oxalates,tartrates and benzoates.

[0067] Salts derived from inorganic or organic bases include alkalimetal salts such as sodium or potassium salts, alkaline earth metalsalts such as magnesium or calcium salts, and organic amine salts suchas morpholine, piperidine, dimethylamine or diethylamine salts.

[0068] Particularly useful salts of compounds according to the inventioninclude pharmaceutically acceptable salts, especially acid additionpharmaceutically acceptable salts.

[0069] In the compounds according to the invention the group R¹ ispreferably an Ar¹L²Ar²Alk-group. In compounds of this type Ar¹ ispreferably an optionally substituted phenyl, monocyclic heteroaromaticor bicyclic heteroaromatic group. Particularly useful monocyclicheteroaromatic groups are optionally substituted five- or six-memberedheteroaromatic groups as described previously, especially five- orsix-membered heteroaromatic groups containing one or two heteroatomsselected from oxygen, sulphur or nitrogen atoms. Nitrogen-containinggroups are especially useful, particularly pyridyl or pyrimidinylgroups. Particularly useful substituents present on these Ar¹ groupsinclude halogen atoms or optionally-substituted alkyl, —OR⁵, —SR⁵,—NR⁵R⁶, —CO₂H, —CO₂CH₃, —NO² or —CN groups as described above inrelation to the compounds of formula (1). Particularly useful bicyclicheteroaromatic groups represented by Ar¹ include optionally substitutedten-membered fused-ring heteroaromatic groups containing one or twoheteroatoms, especially nitrogen atoms. Particular examples includeoptionally substituted naphthyridinyl, especially 2,6-naphthyridinyl,quinolinyl and isoquinolinyl, especially isoquinolin-1-yl groups.Particular optional substituents include those just described formonocyclic heteroaromatic groups.

[0070] A particularly useful group of compounds according to theinvention has the formula (2a):

[0071] wherein —W═ is —CH═ or —N═;

[0072] R¹⁶ and R¹⁷, which may be the same or different is each ahydrogen atom or an atom or group -L³(Alk²)_(t)L⁴(R⁴)_(u) in which L³,Alk², t, L⁴ R⁴ and u are as defined previously;

[0073] L¹, L², Ar², Alk, R², Alk¹, n and R³ are as defined for formula(1);

[0074] and the salts, solvates, hydrates and N-oxides thereof.

[0075] —W═ in compounds of formula (2a) is preferably —N═.

[0076] R¹⁶ and R¹⁷ in compounds of formula (2a) is each preferably asparticularly described above for compounds of formula (1), other than ahydrogen atom. Particularly useful R¹⁶ and R¹⁷ substituents includehalogen atoms, especially fluorine or chlorine atoms, or methyl,halomethyl, especially —CF₃, —CHF₂ or —CH₂F, methoxy or halomethoxy,especially —OCF₃, —OCHF₂ or —OCH₂F groups.

[0077] A further particularly useful group of compounds according to theinvention has the formula (2b):

[0078] wherein R¹⁶, L¹, L², Ar², Alk, R², Alk¹, n and R³ are as definedfor formula (2a);

[0079] g is zero or the integer 1, 2, 3 or 4;

[0080] and the salts, solvates, hydrates and N-oxides thereof.

[0081] Each R¹⁶ atom or group in compounds of formula (2b) may beindependently selected from an atom or group -L³(Alk²)_(t)L³(R⁴)_(u) inwhich L³, Alk², t, L⁴, R⁴ and u are as previously defined. Particularlyuseful R¹⁶ substituents when present in compounds of formula (2b)include halogen atoms; especially fluorine, chlorine or bromine atoms,or methyl, halomethyl, especially —CF₃, methoxy or halomethoxy,especially —OCF₃, —CN, —CO₂Me; —NO₂, amino (—NH₂), substituted amino(—NR⁵R⁶) and —N(R⁵)COCH₃, especially —NHCOCH₃ groups.

[0082] In one preferred group of compounds of formula (2b) each R¹⁶ is ahydrogen atom.

[0083] Another particularly useful group of compounds according to theinvention has the formula (2c):

[0084] wherein R¹⁶, g, L¹, L², Ar², Alk, R², Alk¹, n and R³ are asdefined for formula (2b);

[0085] and the carbon atoms at positions 6 and 7 of the naphthyridinering are indicated with the appropriate numerals;

[0086] and the salts, solvates, hydrates and N-oxides thereof.

[0087] Each R¹⁶ atom or group in compounds of formula (2c) may beindependently selected for an atom or group -L³(Alk²)_(t)L⁴(R⁴)_(u) inwhich L³, Alk², t, L⁴, R⁴ and u are as previously defined. Particularlyuseful R¹⁶ substituents when present in compounds of formula (2c)include halogen atoms, especially fluorine or chlorine atoms, methyl,halomethyl, especially —CF₃, methoxy or halomethoxy, especially —OCF₃,—CN, —CO₂Me, —NO₂, amino (—NH₂), substituted amino (—NR⁵R⁶) and—N(R⁵)COCH₃, especially —NHCOCH₃ groups.

[0088] In one preferred group of compounds of formula (2c) g is theinteger 1 and R¹⁶ is a methoxy group, especially a methoxy group presentat the 6-position. In another preferred group of compounds of formula(2c) g is the integer 2 and each R¹⁶ group is a methoxy group,especially a methoxy group present at the 6- and 7-positions.

[0089] Alk in compounds of the invention is preferably:

[0090] or, especially, —CH₂CH(R)—.

[0091] R in the compounds of formulae (1), (2a), (2b) and (2c) ispreferably a —CO₂H group.

[0092] In general in compounds of formulae (1), (2a), (2b) and (2c) R²is preferably a hydrogen atom.

[0093] In general in compounds of formula (2a) L² is preferably L^(2a)where L^(2a) is a —CON(R⁸)— group, especially —CONH—

[0094] In general in compounds of formulae (2b) and (2c) L² ispreferably L²a where L^(2a) is an —O— atom or —N(R⁸)— group. Anespecially useful —N(R⁸)— group is —NH—.

[0095] The group Ar² in compounds of formulae (1), (2a), (2b) and (2c)is preferably an optionally substituted phenylene group. Particularlyuseful groups include optionally substituted 1,4-phenylene groups.

[0096] In general in compounds of formulae (1), (2a), (2b) and (2c) whenn is zero or the integer 1 the group R³ may especially be a hydrogenatom or an optionally substituted heteroaliphatic, cycloaliphatic,heterocycloaliphatic, aromatic or heteroaromatic group as definedherein. Particularly useful groups of this type include optionallysubstituted C₂₋₆heteroalkyl, particularly C₁₋₃alkoxyC₁₋₃alkyl,especially methoxypropyl, optionally substituted C₃₋₇cycloalkyl,especially optionally substituted cyclopropyl, cyclobutyl, cyclopentyl,cyclopropyl or cyclohexyl, optionally substitutedC₅₋₇heterocycloaliphatic, especially optionally substitutedpyrrolidinyl, piperidinyl or thiazolidinyl, especially optionallysubstituted phenyl and optionally substituted C₅₋₇heteroaromatic,especially optionally substituted pyridyl, pyrimidinyl or triazinylgroups. Optional substituents on these groups include in particular R¹³atoms or groups where the group is an aromatic or heteroaromatic groupand halogen atoms or C₁₋₆alkyl, especially methyl, haloC₁₋₆alkyl,especially trifluoromethyl, C₁₋₆alkoxy, especially methoxy,haloC₁₋₆alkoxy, especially trifluoromethoxy or -(L⁶)_(p)(Alk⁵)_(q)R¹²groups as described earlier where the group is a nitrogen-containingheterocycloaliphatic group such as a pyrrolidinyl, piperidinyl orthiazolidinyl group. Particularly useful -(L⁶)_(p)(Alk⁵)_(q)R¹² groupsinclude those in which L⁶ is a —CO— group. Alk⁵ in these groups ispreferably present (i.e. q is preferably an integer 1) and in particularis a —CH₂— chain.

[0097] Compounds of this type in which R¹² is a hydrogen atom or anoptionally substituted aromatic or heteroaromatic group, especially anoptionally substituted phenyl, pyridyl or imidazolyl group areparticularly preferred.

[0098] In one preferred class of compounds of formulae (1), (2a), (2b)and (2c) L¹ is present as a —N(R⁸)— group. Particularly useful —N(R⁸)—groups include —NH—, —N(CH₃)—, —N(CH₂CH₃)— and —N(CH₂CH₂CH₃)— groups. Inthis class of compounds n is preferably the integer 1 and Alk¹ ispreferably an optionally substituted straight or branched C₁₋₆alkylenechain. Particularly useful Alk¹ chains include —CH₂—, —CH₂CH₂—,—CH₂CH₂CH₂—, —CH(CH₃)CH₂— and —C(CH₃)₂CH₂—. R³ in this class ofcompounds is preferably a hydrogen atom.

[0099] In another preferred class of compounds of formulae (1), (2a),(2b) and (2c) L¹ is a covalent bond, n is the integer 1 and Alk¹ is anoptionally substituted straight or branched C₁₋₆alkylene chain.Particularly useful Alk¹ chains include optionally substituted —CH₂—,—CH₂CH₂—, —CH₂CH₂CH₂— and —CH(CH₃)CH₂— and especially —C(CH₃)₂CH₂—chains. R³ in this class of compounds is preferably a hydrogen atom. Amost especially useful optionally substituted Alk¹ R³ group is —C(CH₃)₃.

[0100] In another preferred class of compounds of fomulae (1), (2a),(2b) and (2c), L¹ is a covalent bond, n is zero and R³ is an optionallysubstituted C₅₋₇heterocycloaliphatic, especially an optionallysubstituted piperidinyl group. A most especially useful optionallysubstituted piperidinyl group is an optionally substitutedpiperidin-1-yl group.

[0101] Particularly useful compounds of the invention include:

[0102](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0103](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-t-butyl-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0104](S)-3-{4-[(6,7-Dimethoxy4-quinazolinyl)amino]phenyl}-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0105](S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0106](S)-3-[4-([6,7-Dimethoxy-4-quinazolinyl)oxy]phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0107](S)-3-[4-([6,7-Methoxy4-quinazolinyl]amino)phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0108](S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-N,N-dipropylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0109](S)-3-[4-([2,6-Naphthyridin-1-yl]oxy)phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0110](S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-piperidin-1-yl-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0111](R)-3-{4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl}-3-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0112](S)-3-[4-([2,6-Naphthyridin-1-yl]oxy)phenyl]-2-[(2-N,N-dipropylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;

[0113] (S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-N,-ethyl-N-isopropylamino-3,4-dioxocyclobut-1-enyl)amino]propanoic acid;

[0114] and the salts, solvates, hydrates and N-oxides thereof.

[0115] The compounds according to the invention are generally of use inmodulating cell adhesion. Thus for example when R¹ in compounds of theinvention is an α₄-integrin binding groups the compounds are of use inthe prophylaxis and treatment of diseases or disorders involvinginflammation in which the extravasation of leukocytes plays a role.

[0116] Diseases or disorders of this type include inflammatory arthritissuch as rheumatoid arthritis vasculitis or polydermatomyositis, multiplesclerosis, allograft rejection, diabetes, inflammatory dermatoses suchas psoriasis or dermatitis, asthma and inflammatory bowel disease.

[0117] In another example when R¹ is an α_(v)-integrin binding group thecompounds may be of use in the prophylaxis and treatment of diseases ordisorders involving inappropriate growth or migration of cells.Particular diseases include inflammatory diseases, and diseasesinvolving angiogenesis, bone resorption or cllular or matrixover-expression.

[0118] Particular uses to which these compounds of the invention may beput include the treatment or inhibition of tumour growth and metastasis;retinopathy; macular degeration psoriasis; rheumatoid arthritis;osteoporosis; bone resorption following or due to joint replacement,hypercalcemia or malignancy, Paget's disease, glucocorticoid treatment,immonilisation-induced osteopenia, hyperparathyroidism or peridontaldisease, vascuar restenosis, atherosclerosis; inflammatory boweldisease; and psoriasis.

[0119] For the prophylaxis or treatment of disease the compoundsaccording to the invention may be administered as pharmaceuticalcompositions, and according to a further aspect of the invention weprovide a pharmaceutical composition which comprises a compound offormula (1) together with one or more pharmaceutically acceptablecarriers, excipients or diluents.

[0120] Pharmaceutical compositions according to the invention may take aform suitable for oral, buccal, parenteral, nasal, topical or rectaladministration, or a form suitable for administration by inhalation orinsufflation.

[0121] For oral administration, the pharmaceutical compositions may takethe form of, for example, tablets, lozenges or capsules prepared byconventional means with pharmaceutically acceptable excipients such asbinding agents (e.g. pregelatinised maize starch, polyvinylpyrrolidoneor hydroxypropyl methylcellulose); fillers (e.g. lactose,microcrystalline cellulose or calcium hydrogen phosphate); lubricants(e.g. magnesium stearate, talc or silica); disintegrants (e.g. potatostarch or sodium glycollate); or wetting agents (e.g. sodium laurylsulphate). The tablets may be coated by methods well known in the art.Liquid preparations for oral administration may take the form of, forexample, solutions, syrups or suspensions, or they may be presented as adry product for constitution with water or other suitable vehicle beforeuse. Such liquid preparations may be prepared by conventional means withpharmaceutically acceptable additives such as suspending agents,emulsifying agents, non-aqueous vehicles and preservatives. Thepreparations may also contain buffer salts, flavouring, colouring andsweetening agents as appropriate.

[0122] Preparations for oral administration may be suitably formulatedto give controlled release of the active compound.

[0123] For buccal administration the compositions may take the form oftablets or lozenges formulated in conventional manner.

[0124] The compounds of formula (1) may be formulated for parenteraladministration by injection e.g. by bolus injection or infusion.Formulations for injection may be presented in unit dosage form, e.g. inglass ampoule or multi dose containers, e.g. glass vials. Thecompositions for injection may take such forms as suspensions, solutionsor emulsions in oily or aqueous vehicles, and may contain formulatoryagents such as suspending, stabilising, preserving and/or dispersingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g. sterile pyrogen-free water,before use. For particle mediated administration the compounds offormula (1) may be coated on particles such as microscopic goldparticles.

[0125] In addition to the formulations described above, the compounds offormula (1) may also be formulated as a depot preparation. Such longacting formulations may be administered by implantation or byintramuscular injection.

[0126] For nasal administration or administration by inhalation, thecompounds for use according to the present invention are convenientlydelivered in the form of an aerosol spray presentation for pressurisedpacks or a nebuliser, with the use of suitable propellant, e.g.dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas ormixture of gases.

[0127] The compositions may, if desired, be presented in a pack ordispenser device which may contain one or more unit dosage formscontaining the active ingredient. The pack or dispensing device may beaccompanied by instructions for administration.

[0128] The quantity of a compound of the invention required for theprophylaxis or treatment of a particular condition will vary dependingon the compound chosen, and the condition of the patient to be treated.In general, however, daily dosages may range from around 100 ng/kg to100 mg/kg e.g. around 0.01 mg/kg to 40 mg/kg body weight for oral orbuccal administration, from around 10 ng/kg to 50 mg/kg body weight forparenteral administration and around 0.05 mg to around 1000 mg e.g.around 0.5 mg to around 1000 mg for nasal administration oradministration by inhalation or insufflation.

[0129] The compounds of the invention may be prepared by a number ofprocesses as generally described below and more specifically in theExamples hereinafter. In the following process description, the symbolsAr¹, Ar², Alk, R¹, R², R³, L¹, L², Alk¹ and n when used in the formulaedepicted are to be understood to represent those groups described abovein relation to formula (1) unless otherwise indicated. In the reactionsdescribed below, it may be necessary to protect reactive functionalgroups, for example hydroxy, amino, thio or carboxy groups, where theseare desired in the final product, to avoid their unwanted participationin the reactions. Conventional protecting groups may be used inaccordance with standard practice [see, for example, Green, T. W. in“Protective Groups in Organic Synthesis”, John Wiley and Sons, 1991]. Insome instances, deprotection may be the final step in the synthesis of acompound of formula (1) and the processes according to the inventiondescribed hereinafter are to be understood to extend to such removal ofprotecting groups. For convenience the processes described below allrefer to a preparation of a compound of formula (1) but clearly thedescription applies equally to the preparation of compounds of formula(2).

[0130] Thus according to a further aspect of the invention, a compoundof formula (1) in which R is a —CO₂H group may be obtained by hydrolysisof an ester of formula (3):

[0131] where Alk represents a group

—CH₂CH(CO₂R^(y))—, —CH═CH(CO₂R^(y))—,

[0132] [where R^(y) is an alkyl group for example a C₁₋₆alkyl group]

[0133] The hydrolysis may be performed using either an acid or a basedepending on the nature of R^(y), for example an organic acid such astrifluoroacetic acid or an inorganic base such as lithium, sodium orpotassium hydroxide optionally in an aqueous organic solvent such as anamide e.g. a substituted amide such as dimethylformamide, an ether e.g.a cyclic ether such as tetrahydrofuran or dioxane or an alcohol e.g.methanol at a temperature from ambient to the reflux temperature. Wheredesired, mixtures of such solvents may be used.

[0134] According to a further aspect of the invention a compound offormula (1) may be prepared by displacement of a leaving group from acompound of formula (4):

[0135] where R^(a) is a leaving group, with an amine R¹R²NH or a saltthereof. Suitable leaving groups represented by R^(a) include halogenatoms, especially chlorine and bromine atoms, or alkoxy, e.g. methoxy,ethoxy or isopropoxy, aryloxy, e g. dinitrophenyloxy, or aralkoxy, e g.benzyloxy, groups.

[0136] The reaction may be performed in an inert solvent or mixture ofsolvents, for example a substituted amide such as dimethylformamide, analcohol such as ethanol and/or a halogenated hydrocarbon such asdichloromethane, at a temperature from 0° C. to the reflux temperature.Where necessary, for example when a salt of an amine R¹, R²NH is used,an organic base such as diisopropylethylamine can be added.

[0137] Any carboxylic acid group present in the intermediate of formula(4) or the amine R¹R²NH may need to be protected during the displacementreaction, for example as an ethyl ester. The desired acid may then beobtained through subsequent hydrolysis, for example as particularlydescribed above and generally described below.

[0138] It will be appreciated that the displacement reaction may also beperformed on a compound of formula (5):

[0139] where R^(b) is a leaving group as defined for R^(a) using anintermediate R³(Alk¹)_(n)L¹H where -L¹H is a functional group such as anamine (—NH₂) using the reaction conditions just described.

[0140] Where desired the displacement reaction may also be performed onan intermediate of formulae (4) or (5), R¹R²NH or R³(Alk¹)_(n)L¹H whichis linked, for example via its R¹ or R³ group, to a solid support, suchas a polystyrene resin. After the reaction the desired compound offormula (1) may be displaced from the support by any convenient method,depending on the original linkage chosen. Intermediates of formulae (4)and (5) are either readily available or may be prepared from anintermediate of formula (6):

[0141] where R^(a) and R^(b) are as previously defined and an amineR¹R²NH or intermediate (R³(Alk¹)_(n)L¹H by displacement as justdescribed for the preparation of compounds of formula (1).

[0142] Intermediates of formulae R¹R²NH and R³(Alk¹)_(n)L¹H may beobtained from simpler, known compounds by one or more standard syntheticmethods employing substitution, oxidation, reduction or cleavagereactions. Particular substitution approaches include conventionalalkylation, arylation, heteroarylation, acylation, thioacylation,halogenation, sulphonylation, nitration, formylation and couplingprocedures. It will be appreciated that these methods may also be usedto obtain or modify other compounds of formulae (1) and (2) whereappropriate functional groups exist in these compounds.

[0143] Thus compounds of the invention and intermediates thereto may beprepared by alkylation, arylation or heteroarylation. For example,compounds containing a -L¹H or -L²H group (where L¹ and L² is each alinker atom or group) may be treated with a coupling agentR³(Alk¹)_(n)X¹ or Ar¹X¹ respectively in which X¹ is a leaving atom orgroup such as a halogen atom, e.g. a fluorine, bromine, iodine orchlorine atom or a sulphonyloxy group such as an alkylsulphonyloxy, e.g.trifluoromethylsulphonyloxy or arylsulphonyloxy, e.g.p-toluenesulphonyloxy group.

[0144] The reaction may be carried out in the presence of a base such asa carbonate, e.g. caesium or potassium carbonate, an alkoxide, e.g.potassium t-butoxide, or a hydride, e.g. sodium hydride, or an organicamine e.g. triethylamine or N,N-diisopropylethylamine or a cyclic amine,such as N-methylmorpholine or pyridine, in a dipolar aprotic solventsuch as an amide, e.g. a substituted amide such as dimethylformamide oran ether, e.g. a cyclic ether such as tetrahydrofuran.

[0145] Intermediates of formula Ar¹X¹ and R³(Alk¹)_(n)X¹ are generallyknown, readily available compounds or may be prepared from knowncompounds by standard substitution and other synthetic procedures, forexample as described herein. Thus for example compounds of formula Ar¹X¹in which, for example, Ar¹ represents a 2,6-naphthyridine group may beprepared from alcohols of formula Ar¹OH by reaction with a halogenatingagent, for example a phosphorous oxyhalide such as phosphorousoxychloride at an elevated temperature e.g. 110° C.

[0146] Intermediate alcohols of formula Ar¹OH in which, for example, Ar¹represents a 2,6-naphthyridine group may be prepared by methods wellknown to a person skilled in the art, e.g. by the method of Sakamoto, T.et al [Chem. Pharm. Bull. 3, 626-633, (1985)].

[0147] Alternatively alkylating agents of formula Ar¹X¹ in which, forexample, Ar¹ represents a 2,6-naphthyridine group may be prepared byreaction of a 2,6-naphthyridine N-oxide or N,N′-dioxide with ahalogenating agent, e.g. a phosphorous oxyhalide such as phosphorousoxychloride to give a 1-halo or 1,5-dihalo-2,6-napthyridinerespectively. In the case of 1,5-dihalo-2,6-napthyridines each halogenatom may be substituted separately by a reagent such as HL²Ar²AlkN(R²)Hor HL³(Alk²)_(t)L⁴(R⁴)_(u) by the particular methods just describedabove.

[0148] 2,6-Napthyridine N-oxides and N,N′-dioxides may be generated fromthe corresponding 2,6-napthyridines group by the general methods ofsynthesis of N-oxides described below or they may be synthesised by themethods of Numata, A. et al (Synthesis, 1999, 306-311).

[0149] Further alkylating agents of formula Ar¹X¹ in which, for example,Ar¹ represents a 2,6-naphthyridine, may be prepared by the methods ofGiacomello G. et al (Tetrahedron Letters 1965, 1117-1121), Tan, R. andTaurins, A. (Tetrahedron Letters 1965, 2737-2744), Ames, D. E. andDodds, W. D. (J. Chem. Soc. Perkin 1 1972. 705-710) and Alhaique, F. etal (Tetdrahedron Letters, 1975, 173-174).

[0150] In a further example intermediates of formula R¹, R²NH may beobtained by reaction of a compound of formula Ar¹ L²H with a compound offormula X¹ Ar²AlkN(R²)H under the reaction conditions just described

[0151] Compounds of formula Ar¹L²H in which, for example Ar¹ representsa 2,6-naphthyridine and L² is a —N(R⁸)— group, may be prepared fromsubstituted 4-cyano-3-cyanomethylpyridines by the methods of Alhaique,F. et al (ibid and Gazz. Chim. Ital. 1975, 105, 1001-1009) or from3-fomylpyridines by the methods of Molina, P. at al (Tetrahedron 1992,48, 4601-4616).

[0152] In another example, compounds containing a -L¹H or -L²H or groupas defined above may be functionalised by acylation or thioacylation,for example by reaction with one of the alkylating agents just describedbut in which X¹ is replaced by a —C(O)X², C(S)X², —N(R⁸)COX² or—N(R⁸)C(S)X² group in which X² is a leaving atom or group as describedfor X¹. The reaction may be performed in the presence of a base, such asa hydride, e.g. sodium hydride or an amine, e.g. triethylamine orN-methylmorpholine, in a solvent such as a halogenated hydrocarbon, e.g.dichloromethane or carbon tetrachloride or an amide, e.g.dimethylformamide, at for example ambient temperature. Alternatively,the acylation may be carried out under the same conditions with an acid(for example one of the alkylating agents described above in which X¹ isreplaced by a —CO₂H group) in the presence of a condensing agent, forexample a diimide such as 1-(3-dimethylaminopropyl)-3-ethylcarbodiimideor N,N′-dicyclohexylcarbodiimide, advantageously in the presence of acatalyst such as a N-hydroxy compound e.g. a N-hydroxytriazole such as1-hydroxybenzotriazole. Alternatively the acid may be reacted with achloroformate, for example ethylchloroformate, prior to the desiredacylation reaction

[0153] In a further example compounds may be obtained by sulphonylationof a compound containing an —OH group by reaction with one of the abovealkylating agents but in which X¹ is replaced by a —S(O)Hal or —SO₂Halgroup in which Hal is a halogen atom such as chlorine atom] in thepresence of a base, for example an inorganic base such as sodium hydridein a solvent such as an amide, e.g. a substituted amide such asdimethylformamide at for example ambient temperature.

[0154] In another example, compounds containing a -L¹H or -L²H group asdefined above may be coupled with one of the alkylation agents justdescribed but in which X¹ is replaced by an —OH group in a solvent suchas tetrahydrofuran in the presence of a phosphine, e.g.triphenylphosphine and an activator such as diethyl, diisopropyl- ordimethylazodicarboxylate.

[0155] In a further example, ester groups —CO₂R⁵, —CO₂Alk³ or —CO₂Alk⁷in the compounds may be converted to the corresponding acid [-CO₂H] byacid- or base-catalysed hydrolysis depending on the nature of the groupsR⁵, Alk³ or Alk⁷. Acid- or base-catalysed hydrolysis may be achieved forexample by treatment with an organic or inorganic acid, e.g.trifluoroacetic acid in an aqueous solvent or a mineral acid such ashydrochloric acid in a solvent such as dioxan or an alkali metalhydroxide, e.g. lithium hydroxide in an aqueous alcohol, e.g. aqueousmethanol.

[0156] In a further example, —OR⁵ or —OR¹⁴ groups [where R⁵ or R¹⁴ eachrepresents an alkyl group such as methyl group] in compounds of formula(1) may be cleaved to the corresponding alcohol —OH by reaction withboron tribromide in a solvent such as a halogenated hydrocarbon, e.g.dichloromethane at a low temperature, e.g. around −78° C.

[0157] Alcohol [—OH] groups may also be obtained by hydrogenation of acorresponding —OCH₂R¹⁴ group (where R¹⁴ is an aryl group) using a metalcatalyst, for example palladium on a support such as carbon in a solventsuch as ethanol in the presence of ammonium formate, cyclohexadiene orhydrogen, from around ambient to the reflux temperature. In anotherexample, —OH groups may be generated from the corresponding ester[CO₂Alk⁵ or CO₂R⁵] or aldehyde [—CHO] by reduction, using for example acomplex metal hydride such as lithium aluminium hydride or sodiumborohydride in a solvent such as methanol.

[0158] In another example, alcohol —OH groups in the compounds may beconverted to a corresponding —OR⁵ or —OR¹⁴ group by coupling with areagent R⁵OH or R¹⁴0H in a solvent such as tetrahydrofuran in thepresence of a phosphine, e.g. triphenylphosphine and an activator suchas diethyl-, diisopropyl-, or dimethylazodicarboxylate.

[0159] Aminosulphonylamino [—NHSO₂NHR³ or —NHSO₂NHAr¹] groups in thecompounds may be obtained, in another example, by reaction of acorresponding amine [—NH₂] with a sulphamide R³NHSO₂NH₂ or Ar¹ NHSO₂NH₂in the presence of an organic base such as pyridine at an elevatedtemperature, e.g. the reflux temperature.

[0160] In another example compounds containing a —NHCSAr¹, —CSNHAr¹,—NHCSR³ or —CSNHR³ may be prepared by treating a corrsponding compoundcontaining a —NHCOAr^(a), —CONHAr¹, —NHCOR³ or —CONHR³ group with athiation reagent, such as Lawesson's Reagent, in an anhydrous solvent,for example a cyclic ether such as tetrahydrofuran, at an elevatedtemperature such as the reflux temperature.

[0161] In a further example amine (—NH₂) groups may be alkylated using areductive alkylation process employing an aldehyde and a borohydride,for example sodium triacetoxyborohyride or sodium cyanoborohydride, in asolvent such as a halogenated hydrocarbon, e.g. dichloromethane, aketone such as acetone, or an alcohol, e.g. ethanol, where necessary inthe presence of an acid such as acetic acid at around ambienttemperature.

[0162] In a further example, amine [—NH₂] groups in compounds of formula(1) may be obtained by hydrolysis from a corresponding imide by reactionwith hydrazine in a solvent such as an alcohol, e.g. ethanol at ambienttemperature.

[0163] In another example, a nitro [—NO₂] group may be reduced to anamine [—NH₂], for example by catalytic hydrogenation using for examplehydrogen in the presence of a metal catalyst, for example palladium on asupport such as carbon in a solvent such as an ether, e.g.tetrahydrofuran or an alcohol e.g. methanol, or by chemical reductionusing for example a metal, e.g. tin or iron, in the presence of an acidsuch as hydrochloric acid.

[0164] Aromatic halogen substituents in the compounds may be subjectedto halogen-metal exchange with a base, for example a lithium base suchas n-butyl or t-butyl lithium, optionally at a low temperature, e.g.around −78° C., in a solvent such as tetrahydrofuran and then quenchedwith an electrophile to introduce a desired substituent. Thus, forexample, a formyl group may be introduced by using dimethylformamide asthe electrophile; a thiomethyl group may be introduced by usingdimethyldisulphide as the electrophile.

[0165] In another example, sulphur atoms in the compounds, for examplewhen present in a linker group L¹ or L² may be oxidised to thecorresponding sulphoxide or sulphone using an oxidising agent such as aperoxy acid, e.g. 3-chloroperoxybenzoic acid, in an inert solvent suchas a halogenated hydrocarbon, e.g. dichloromethane, at around ambienttemperature.

[0166] In another example compounds of formula Ar¹X¹ (where X¹ is ahalogen atom such as a chlorine, bromine or iodine atom) may beconverted to such compounds as Ar¹CO₂R²⁰ (in which R²⁰ is an optionallysubstituted alkyl, aryl or heteroaryl group), Ar¹CHO, Ar¹CHCHR²⁰,Ar¹CCR²⁰, Ar¹N(R²⁰)H, Ar¹N(R²⁰)₂, for use in the synthesis of forexample compounds of formula R¹R²NH, using such well known and commonlyused palladium mediated reaction conditions as are to be found in thegeneral reference texts Encyclopedia of Reagents for Organic Synthesis,Editor-in Chief Paquette, L. A., John Wiley and Sons, 1995 andComprehensive Organic Functional Group Transformations, Editors-in-ChiefKatritzky, A R. et al, Pergamon, 1995.

[0167] N-oxides of compounds of formula (1) may be prepared for exampleby oxidation of the corresponding nitrogen base using an oxidising agentsuch as hydrogen peroxide in the presence of an acid such as aceticacid, at an elevated temperature, for example around 70° C. to 80° C, oralternatively by reaction with a peracid such as peracetic acid in asolvent, e.g. dichloromethane, at ambient temperature.

[0168] Salts of compounds of formula (1) may be prepared by reaction ofa compound of formula (1) with an appropriate base in a suitable solventor mixture of solvents e.g. an organic solvent such as an ether e.g.diethylether, or an alcohol, e.g. ethanol using conventional procedures.

[0169] Where it is desired to obtain a particular enantiomer of acompound of formula (1) this may be produced from a correspondingmixture of enantiomers using any suitable conventional procedure forresolving enantiomers.

[0170] Thus for example diastereomeric derivatives, e.g. salts, may beproduced by reaction of a mixture of enantiomers of formula (1) e.g. aracemate, and an appropriate chiral compound, e.g. a chiral base. Thediastereomers may then be separated by any convenient means, for exampleby crystallisation and the desired enantiomer recovered, e.g. bytreatment with an acid in the instance where the diastereomer is a salt.

[0171] In another resolution process a racemate of formula (1) may beseparated using chiral High Performance Liquid Chromatography.Alternatively, if desired a particular enantiomer may be obtained byusing an appropriate chiral intermediate in one of the processesdescribed above.

[0172] Chromatography, recrystallisation and other conventionalseparation procedures may also be used with intermediates or finalproducts where it is desired to obtain a particular geometric isomer ofthe invention.

[0173] The following Examples illustrate the invention. All temperaturesare in ° C. The following abbreviations are used: NMMN-methylmorpholine; EtOAc ethyl acetate; MeOH methanol; BOCbutoxycarbonyl; DCM dichloromethane; AcOH acetic acid; DIPEAdiisopropylethylamine; EtOH ethanol; Pyr pyridine; Ar aryl; DMSOdimethylsulphoxide; iPr isopropyl; Et₂O diethylether; Me methyl; THFtetrahydrofuran, DMF N,N-dimethylformamide; FMOC 9-fluorenylmethoxy- brbroad; carbonyl; obs obscured; app apparent; dil dilute; RT roomtemperature; Bu butyl; DIPEA diisopropylethylamine

[0174] All NMR's were obtained at 300 mHz.

[0175] Intermediate 1

[0176] 3,5-Dichloropyridine-4-carboxylic acid

[0177] A solution of 3,5-dichloropyridine (5.00 g, 33.8 mmol) in THF (25ml) was added to a solution of LDA [generated from nBuLi (2.5M solutionin hexanes, 14.9 ml, 37.2 mmol) and diisopropylamine (4.10 g, 5.7 ml,40.6 mmol)] in THF (25 ml) at −78° under nitrogen, to give ayellow/brown slurry. The reaction was stirred for 30 min at −780 thenCO₂ gas was bubbled through to give a clear brown solution that slowlygave a precipitate, warmed to RT over 2 h, then quenched with water (20ml) and partitioned between Et₂O (100 ml) and 1M NaOH (100 ml). Theaqueous layer was separated and acidified to pH 1 with concentratedhydrochloric acid and then extracted with 10% MeOH in DCM (100 ml×3).The combined organic layers were dried (MgSO₄) and the solvent removedunder vacuum to give a brown solid that was recrystallised from ethanoland dried under vacuum to give the title compound as pinkish crystals(2.63 g, 41%). δ_(H) (DMSO-d⁶) 8.74 (2H, s). δC (DMSO-d⁶) 163.5, 147.7,141.0, 126.7

[0178] Intermediate 2

[0179] Ethyl(S)-3-(4-[3,5-dichloropyrid4-ylcarboxamido]phenyl)-2-(t-butoxycarbonylamino)propionate

[0180] A slurry of the compound of Intermediate 1 (51.2 g, 0.267 mol) inDCM (195 ml) and thionyl chloride (195 ml, 2.67 mol) was treated withDMF (5 drops) and heated to reflux for 4 h. The reaction wasconcentrated in vacuo and azeotroped with toluene (2×50 ml) to give ayellow solid which was used without further purification. A solution ofethyl-(S)-3-(4-aminophenyl)-2-(t-butoxycarbonyl amino)propionate (130.8g, 0.425 mol) in DCM (800 ml) was cooled to 0° and treated with NMM(56.0 ml, 0.51 mol), stirred 5 minutes and then a solution of the acidchloride (98.3 g, 0.468 mol) in DCM (200 ml) was added dropwise keepingthe reaction temperature below 50. The reaction was stirred for 1 h,quenched with NaHCO₃ solution (500 ml), the organic layer separated,washed with NaHCO₃ solution (500 ml), 10% citric acid solution (500 ml)and NaHCO₃ solution (500 ml), dried (MgSO₄)and concentrated in vacuo togive a yellow solid which was recrystallised (EtOAc/hexane) to give thetitle compound, 140 g, 69%. δ_(H) (DMSO d6), 8.8 (2H, s), 7.55 (2H, d, J8.5 Hz), 7.23 (2H, d, J 8.5 Hz), 4.0 (3H, m), 3.4 (2H, b s), 2.9 (1H,m), 2.8 (1H, m), 1.3 (9H, s), 1.25 (3H, t). m/z (ES⁺, 70V) 504 (MNa⁺).

[0181] Intermediate 3

[0182] Ethyl (S)-3-[4(3,5-dichloropyrid4-yl carboxamido)phenyl]-2-aminopropionate hydrochloride

[0183] A solution of the compound of Intermediate 2 (70 g, 0.146 mol) inEtOAc (500 ml) and 1,4-dioxan (50 ml) was treated with a solution of HClin EtOAc (500 ml, 3M), and stirred at RT for 4 h. The reaction wasconcentrated in vacuo to give a yellow solid which was triturated withEt₂O then recrystallised (EtOAc/hexane) to give the title compound (59.3g, 92%). δ_(H) (DMSO d⁶), 11.10 (1H, s), 8.70 (2H, s), 7.55 (2H, d, J8.4 Hz), 7.25 (2H, d, J 8.4 Hz), 4.10 (3H, m), 3.10 (2H, m), 1.10 (3H,m). m/z (ES⁺, 70V) 382 (MH⁺).

[0184] Intermediate 4

[0185] 3-(tert-Butyl)-4-isopropoxy-3-cyclobutene-1,2-dione tert-Butyllithium (2.29 ml of a 1.7M solution in pentane, 3.9 mmol) was added to asolution of 3,4-diisopropoxy-3-cyclobutene-1,2-dione (594 mg, 3 mmol) inTHF (30 ml) at −78° C. After 5 h trifluoroactic anhydride (636 μl, 4.5mmol) was added and stirring continued at −78° C. for 30 min. The coldmixture was poured into NH₄Cl(aq), extraced with EtOAc, dried (Na₂SO₄)and evaporated in vacuo. Column chromatography (S)O₂; EtOAc/hexane,15:85) gave the title compound as a mobile yellow oil (408 mg, 69%). 5H(CDCl₃) 5.43 (1H, sept, 16.2 Hz), 1.45 (6H, d, 16.2 Hz) and 1.33 (9H,s); m/z (ES⁺, 70V) 197 (M⁺+H).

[0186] Intermediate 5

[0187]1-Chloro-2,6-naphthyridine

[0188] 1-Hydroxy-2,6-naphthyridine (550 mg) [prepared according to themethod of Sakamoto, T. et al Chem. Pharm. Bull. 33, 626, (1985)] wasstirred with phosphorous oxychloride (10 ml) at 110° for 5 h. Thevolatiles were removed in vacuo and the residue treated carefully withice. After diluting with water (to ˜25 ml), solid NaHCO₃ was added toneutralise and the product extracted into EtOAc (2×80 ml). The combinedorganic extracts were dried (MgSO₄), evaporated in vacua, and the crudeproduct chromatographed (S)O₂; EtOAc) affording the title compound as aslightly yellow solid (420 mg, 68%). δ_(H) (CDCl₃) 9.35 (1H, s), 8.82(1H, d, J 5.9 Hz), 8.48 (1H, d, 15.6 Hz), 8.00 (1H, d, 15.9 Hz), 7.74(1H, d, J 5.6 Hz); m/z (ES⁺, 70V) 165 and 167 (MH⁺).

[0189] Intermediate 6

[0190] Ethyl(S)-3-{4-[(2,6-naphthyridin-1-yl)amino]phenyl}-2-[N-(t-butyloxycarbonyl) amino]propanoate

[0191] Ethyl(S)-3-(4-aminophenyl)-2-[N-(t-butyloxycarbonyl)amino]propanoate (600 mg,1.95 mmol), Intermediate 5 (350 mg, 2.13 mmol) and DIPEA (276 mg, 372μl, 2.13 mmol) in 2-ethoxyethanol (0.5 ml) were stirred at 130° under N₂for several hours. The reaction was partitioned between EtOAc (70 ml)and saturated aqueous NaHCO₃ (30 ml). The phases were separated and theaqueous layer re-extracted with EtOAc (3×30 ml). The combined organicextracts were washed with brine (10 ml), dried (MgSO₄) and evaporated invacuo to afford a dark oil. Chromatography (S)O₂; 3% MeOH/DCM) gave thetitle compound as a dull orange foam (360 mg, 42%). δ_(H) (CDCl₃) 9.19(1H, s), 8.67 (1H, d, J 5.9 Hz), 8.24 (1H, d, J 5.8 Hz), 7.66 (1H, d, J5.9 Hz), 7.65 (2H, d, J 8.5 Hz), 7.21 (1H, d, J 5.8 Hz), 7.16 (2H, d, J8.5 Hz), 7.15 (1H, obscured s), 5.05-4.97 (1H, m), 4.60-4.51 (1H, m),4.19 (2H, q, 17.1 Hz), 3.17-3.04 (2H, m), 1.44 (9H, s), 1.27 (3H, t, J7.1 Hz); m/z (ES⁺, 70V) 459 (MNa⁺), 437 (MH⁺).

[0192] Intermediate 7

[0193] Ethyl(S)-2-amino-3-{4-[(2,6-naphthyridin-1-yl)amino]phenyl}propanoate

[0194] Intermediate 6 (360 mg) was treated with a solution oftrifluoroacetic acid (10 ml) and DCM (10 ml) and stirred at RT for 2 h.The volatiles were removed in vacuo and the residue was partitionedbetween EtOAc (80 ml) and saturated aqueous NaHCO₃ (30 ml). The phaseswere separated and the aqueous layer re-extracted with EtOAc (3×30 ml).The combined organic extracts were dried (MgSO₄) and evaporated in vacuoto afford the title compound as a dark orange viscous oil (280 mg,100%). δ_(H) (CDCl₃) 9.18 (1H, s), 8.66 (1H, d, J 5.9 Hz), 8.22 (1H, d,J 5.8 Hz), 7.67 (1H, d, J 5.9 Hz), 7.64 (2H, d, J 8.5 Hz), 7.22 (2H, d,J 8.5 Hz), 7.19 (1H, d, J 5.8 Hz), 4.20 (2H, q, J 7.1 Hz), 3.73 (1H, dd,J 7.9, 5.1 Hz), 3.10 (1H, dd, J 13.6, 5.2 Hz), 2.87 (1H, dd, J 13.6, 7.9Hz), 1.70 (3H, br s), 1.28 (3H, t, 7.1 Hz); m/z (ES⁺, 70V) 337 (MH⁺).

[0195] Intermediate 8

[0196] Methyl(S)-2-(t-butyloxycarbonylamino)-3-[4-(2,6-naphthyridin-1-yloxy)phenyl]propanoate

[0197] To N-(t-butyloxycarbonyl) tyrosine methyl ester (1.42 g, 4.82mmol) in dry DMF (10 ml) was added 1-chloro-2,6 naphthyridine (0.79 g,4.82 mmol) and cesium carbonate (1.65 g, 5.06 mmol) and the reactionstirred at 450 under N₂ for 2 days. The DMF was evaporated, EtOAc addedand washed (3×) with water, dried (MgSO₄), and evaporated in vacuo. Theresidue was chromatographed (SiO₂; 40 to 100% EtOAc/isohexane) to affordthe tile compound as white foam (1.61 g, 82%). δ_(H) (CDCl₃) 9.29 (1H,s), 8.76 (1H, d, J 5.74 HZ), 8.17 (1H, d, J 5.74 Hz), 8.11 (1H, d, J 5.8Hz), 7.43 (1H, d, J 5.8 Hz), 7.22-7.18 (3H, m), 5.03 (1H, br s), 4.61(1H, br s), 3.75 (3H, s), 3.15-3.05 (2H, m), 1.44 (9H, s); m/z (ES⁺,70V) MH⁺ 424.

[0198] Intermediate 9

[0199] Ethyl(S)-2-(N-t-butyloxycarbonylamino)-3-[4-(isoquinolin-1-ylamino)phenyl]propanoate

[0200] A stirred solution of ethyl(S)-3-(4-aminophenyl)-2-(N-4-butyloxycarbonylamino)propanoate (3.08 g,10.0 mmol), 1-chloroisoquinoline (1.80 g, 11.0 mmol) andN,N-diisopropylethylamine (1.42 g, 1.91 ml, 11.0 mmol) in2-ethoxyethanol (1.0 ml) was heated at 130° for 4 h. The volatiles wereremoved in vacuo and the residue partitioned between EtOAc (120 ml) andsaturated aqueous NaHCO₃ (50 ml). The phases were separated and theaqueous layer was re-extracted with EtOAc (80 ml). The combined organicextracts were washed with brine (30 ml), dried (MgSO₄) and evaporated invacuo. The obtained dark oil was chromatographed (Silica; 20-30%EtOAc/hexane) to afford the title compound as a pink oil whichcrystallised on standing (2.78 g, 64%). δ_(H) (CDCl₃) 8.07 (1H, d, J 5.8Hz), 7.93 (1H, d, J 8.4 Hz), 7.72 (1H, d, J 7.5 Hz), 763 (1H, d), 7.61(2H, d, B 8.5 Hz), 7.51 (1H, t, J 6.8 Hz), 7.23 (1H, br s), 7.10 (1H, brs), 7.10 (2H, d, J 6.8 Hz), 5.02 (1H, br d, J 8.0 Hz), 4.54 (1H, br m),4.16 (2H, t, J 7.1 Hz), 3.05 (2H, br m), 1.43 (9H, s), 1.25 (3H, t, J7.1 Hz); m/z (ES⁺, 60V) 436 (MH⁺).

[0201] Intermediate 10

[0202] (S)-Ethyl 2-amino-3-[4-(isoquinolin-1-ylamino)phenyl]propanoate

[0203] A stirred solution of Intermediate 9 (2.709) in EtOAc (100 ml)was treated with HCl gas until turbidity and precipitation was seen tooccur. The reaction mixture was stirred at ambient temperatue for anaddition 0.5 h. The reaction was purged with nitrogen then diluted withEtOAc (50 ml) and saturated aqueous NaHCO₃ (50 ml). Sufficient solidNaHCO₃ was added to ensure full neutrality. The phases were separatedand the aqueous layer re-extracted with EtOAc (2×40 ml). The combinedorganic extracts were washed with brine (20 ml), dried (MgSO₄) andevaporated in vacuo to afford the title compound as a light orange oil(2.10 g, q). δH (CDCl₃) 8.06 (1H, d, J 5.8 Hz), 7.91 (1H, d, 18.3 Hz),7.71 (1H, d, J 7.9 Hz), 7.63 (1H, obs. signal), 7.59 (2H, d, J 8.4 Hz),7.49 (1H, app.t, J 7.8 HZ), 7.25 (1H, br s), 7.15 (1H, d, J 8.4 Hz),7.09 (1H, d, J 5.8 Hz), 4.17 (2H, q, J 7.2 Hz), 3.68 (1H, dd, J 7.7, 5.1Hz), 3.06 (1H, dd, J 14.6, 5.1 Hz), 2.81 (1H, dd, J 13.6, 7.9 Hz), 1.58(2H, br s), 1.26 (3H, t, v7.0 Hz); m/z (ES⁺, 60V) 435.9 (MH⁺).

[0204] Intermediate 11

[0205] Ethyl (E)-3-{4-[(tert-Butoxycarbonyl)amino]phenyl}-2-propenoate

[0206] Ethyl 4-aminocinnamate (2.5 g, 13.1 mmol) was dissolved in THF(25 ml) and treated with di-tert-butyl dicarbonate (3.14 g). Thesolution was refluxed for 16 h and then allowed to cool. The product wasextracted into EtOAc and washed with water and brine, dried over Na₂SO₄,filtered and the solvent removed. The crude product was purified bycolumn chromatography (S)O₂; EtOAc/hexane 1:9) to give the titlecompound (2.82 g, 74%) as a white solid. δ_(H) (CDCl₃) 7.62 (1H, d, J16.0 Hz), 7.45 (2H, d, 8.8 Hz), 7.38 (2H, d, 18.8 Hz), 6.63 (1H, br s),6.33 (1H, d, J 16.0 Hz), 4.12 (2H, q, J 7.1 Hz), 1.52 (9H, s), 1.25 (3H,t, J 7.1 Hz). m/z (ES⁺, 70V) 314 (MNa⁺).

[0207] Intermediate 12

[0208] Ethyl(3S)-3-{4-[(tert-Butoxycarbonyl)amino]phenyl}3-{N-benzyl[(1R)-1-phenylethyl]amino}propanoate

[0209] Intermediate 11 (1.0 g, 3.44 mmol) was dissolved in THF (25 ml),treated with sodium hydride and left to stir for 20 mins.(R)-(+)—N-Benzyl-(methylbenzylamine (1.44 ml) in THF (25 ml) at 0° wastreated with n-butyllithium (2.75 ml, 2.5M in hexanes) and the purplesolution left to stir for 20 mins then cooled to −78° and the esteranion added slowly. The reaction mixture was stirred at −78° C. for 4 hthen quenched with ammonium chloride solution, extracted into EtOAc,washed with water and brine, dried (Na₂SO₄), filtered and the solventremoved. The crude product was purified by column chromatography (S)O₂;CH₂Cl₂) to give the title compound (1.14 g, 66%) as a white solid. δ_(H)(CDCl₃) 7.42-7.17 (14H, m), 6.45 (1H, br s), 4.39 (1H, dd, J 9.4, 5.5Hz), 3.99 (1H, q, J 6.9 Hz), 3.93 (2H, qd, J 7.1, 2.4 Hz), 3.72 (1H, d,J 14.7 Hz), 3.64 (1H, d, J 14.7 Hz), 2.63 (1H, dd, J 14.7, 5.5 Hz), 2.52(1H, dd, J 14.7, 9.5 Hz), 1.52 (9H, s), 1.22 (3H, d, J 6.9 Hz), 1.06(3H, t, 17.1 Hz). m/z (ES⁺, 70V) 503 (MH⁺).

[0210] Intermediate 13

[0211] Ethyl(3S)-3-amino-3-{4-[(tert-Butoxycarbonyl)amino]}phenyl-propanoate

[0212] Intermediate 12 (312 mg, 0.62 mmol) in MeOH (5 ml) was treatedwith formic acid (0.1 ml) and 10% palladium on carbon. The reactionmixture was heated to reflux for 30 mins then cooled, filtered throughcelite™ and the solvent removed to give the title compound (195 mg,100%) as an oil. δ_(H) (CDCl₃) 8.05 (2H, br s), 7.28 (2H, d, J 8.5 Hz),7.21 (2H, d, J 8.5 Hz,), 6.92 (1H, brs), 4.48 (1H, dd, J 8.4, 5.7 Hz),4.05 (2H, q, J 7.1 Hz), 3.6 (1H, dd, J 17.2, 8.4 Hz), 2.79 (1H, dd, J17.2, 5.7 Hz), 1.44 (9H, s), 1.14 (3H, t, J 7.1 Hz). m/z (ES⁺, 70V) 331(MNa⁺).

[0213] Intermediate 14

[0214] Methyl(R)-3-[(tert-Butoxycarbonyl)amino]-3-(4-hydroxyphenyl)propanoate

[0215] Methyl (3R)-(3-amino)-3-(4-hydroxyphenyl)propanoate [S. G. Daviesand 0. Ichihara, Tetrahedron Asymmetry, (1991), 2, 183-186] (346 mg,1.78 mmol) was dissolved in dioxan (5 ml) and sodium bicarbonatesolution (5 ml) added. The solution was treated with di-tert-butyldicarbonate (407 mg, 1.86 mmol) and stirred vigourously for 16 h. Thesolution was diluted with water, and the product extracted into EtOAc(×2), washed with water, brine, dried (Na₂SO₄), filtered and the solventremoved. The product was purified by column chromatography (S)O₂;CH₂Cl₂/MeOH 20:1) to give the title compound (211 mg, 42%) as a whitesolid. δ_(H) (CDCl₃) 7.06 (2H, d, J 8.6 Hz), 6.66 (2H, d, J 8.6 Hz),5.48 (1H, br), 4.98 (2H, br m), 3.61 (3H, s), 2.78 (2H, m), 1.42 (9H,s). m/z (ES⁺, 70V) 318 (MNa⁺).

[0216] Intermediate 15

[0217] Methyl(3R)-3-[(tert-Butoxycarbonyl)amino]-3-{4-[(6,7-dimethoxy-4-quinazolinyl)oxy]phenyl}propanoate

[0218] Intermediate 14 (420 mg, 1.42 mmol) in DMF (4 ml) was treatedwith potassium carbonate (394 mg) and 4-chloro-6,7-dimethoxyquinazoline(320 mg). The solution was stirred for 48 h and then water (20 ml) wasadded. The mixture was extracted with EtOAc (×2), washed with water(×3), brine, dried (Na₂SO₄), filtered and the solvent removed to givethe title compound (657 mg, 96%) as a foamy yellow solid. δ_(H) (DMSOd⁶) 8.53 (1H, s), 7.53 (1H, s), 7.40 (2H, d, J 8.6 Hz), 7.37 (1H, s),7.24 (2H, d, J 8.6 Hz), 4.96 (1H;, m, CH), 3.98 (3H, s), 3.95 (3H, s),3.57 (3H, s), 2.77 (2H, m), 1.36 (9H, s). m/z (ES⁺, 70V) 484 (MH⁺).

[0219] Intermediate 16

[0220] Methyl(3R)-3-Amino-3-{4-[(6,7-dimethoxy4-quinazolinyl)oxy]phenyl}propanoate

[0221] Intermediate 15 (650 mg, 1.35 mmol) was dissolved in EtOAc (10ml) and HCl gas was bubbled through. The reaction mixture was stirredfor 2 h and the solvent removed to give the title compound (589 mg,100%) as an oil. δ_(H) (DMSO d⁶) 8.66 (1H, s), 7.65 (2H, d, J 8.7 Hz),7.58 (1H, s), 7.44 (1H, s), 7.39 (2H, d, J 8.7 Hz), 3.99 (3H, s), 3.97(3H, s), 3.58 (3H, s), 3.22 (1H, dd, J 16.3, 6.1 Hz), 3.05 (1H, dd, J16.3, 8.5 Hz). m/z (ES⁺, 70V) 384 (MH⁺).

[0222] Intermediate 17

[0223] Methyl (S)-3-{4-[(3-phenyl-1quinazolinyl)amino]-phenyl}-[2-(tert-butoxycarbonyl)amino]-propanoate

[0224] Methyl(2S)-[2-(tert-butoxycarbonyl)amino]-3-(4-aminophenyl)propanoate (500 mg,1.7 mmol) and 4-chloro-2-phenylquinazoline (408 mg) were dissolved in2-ethoxyethanol (5 ml) with Hunigs base (0.6 ml) and the solution heatedat 120° C. for 16 h. The solution was cooled and concentrated. Theresidue was purified by column chromatography (S)O₂; CH₂Cl₂/MeOH 25:1)to give the title compound (682 mg, 81%) as a brown foamy solid. δ_(H)(CDCl₃) 8.56 (2H, dd, J 7.5, 3.7 Hz), 8.10 (1H, m), 7.95 (1H, m), 7.88(2H, d, J 8.5 Hz), 7.80 (1H, m), 7.70 (1H, m), 7.50 (3H, m), 7.23 (2H,d, J 8.5 Hz), 5.05 (1H, m), 4.65 (1H, m), 3.72 (3H, s), 3.49 (1H, m),3.15 (2H, m), 1.45 (9H, s). m/z (ES⁺, 70V) 499 (MH⁺).

[0225] Intermediate 18

[0226] Methyl (S)-2-Amino-3-{4-[(3-phenyl-1-quinazolinylamino]phenyl}propanoate

[0227] Intermediate 17 (678 mg, 1.36 mmol) in EtOAc (30 ml) wassaturated with HCl gas and stirred for 45 mins. The brown precipitatewas filtered off and dried to give the title compound (518 mg, 96%) as abrown foamy solid. 5H (DMSO d⁶) 9.12 (1H, d, J 8.6 Hz), 8.83 (2H, m),8.50 (1H, d, J 8.1 Hz), 8.44 (2H, d, J 7.1 Hz), 8.14 (1H, d, J 8.1 Hz),8.10 (1H, t, J 8.1 Hz), 7.84 (2H, d, u 8.6 Hz), 7.70 (1H, d, J 7.1 Hz),7.63 (2H, t, J 17.1 Hz), 7.40 (2H, d, J 8.5 Hz), 3.70 (3H, s), 3.67 (1H,m), 3.30 (1H, dd, J 14.0, 5.5 Hz), 3.18 (1H, dd, J 14.0, 7.4 Hz). m/z(ES⁺, 70V) 399 (MH⁺).

[0228] Intermediate 19

[0229] Ethyl(R)-3-Amino-3-[4-(tert-butoxycarbonyl)aminophenyl]propanoate

[0230] Ethyl(3R)-3-{Benzyl[(1S)-1-phenylethyl]amino}-3-[4-(tert-butoxycarbonyl)amino phenyl]propanoate (1.18 g, 2.35 mmol) was dissolved in MeOH (10ml) and formic acid (1 ml) and 10% palladium on carbon added and themixture refluxed for 2 h. The reaction mixture was cooled, filteredthrough Celite® and concentrated to give the crude title compound whichwas used immediately in the next reaction. δ_(H) (CDCl₃) 7.34 (2H, d, J8.1 Hz), 7.27 (2H, d, J 8.1 Hz), 7.10 (1H, br s), 4.59 (1H, m), 4.11(2H, q, J 7.1 Hz), 3.14 (1H, dd, J 16.8, 7.9 Hz), 2.86 (1H, dd, J 16.8,12.0 Hz), 1.20 (3H, t, J 7.1 Hz).

[0231] Intermediate 20

[0232] Ethyl (R)-3-amino-3-(4-aminophenyl)propanoate Intermediate 19 wasdissolved in EtOAc (25 ml) and the solution saturated with HCl gas. Thesolution was stirred at RT for 90 mins whilst a white precipitateformed. The solid was filtered and dried to give the title compound (570mg, 88% over 2 steps) as a white solid. AH (DMSO d6) 8.79 (2H, br s),7.63 (2H, d, J 8.4 Hz), 7.36 (2H, d, J 8.4 Hz, 4.58 (1H, m), 3.98 (2H,q, J 7.1 Hz), 3.19 (1H, dd, J 16.3, 5.6 Hz), 2.99 (1H, dd, J 16.3, 9.1Hz), 1.08 (3H, t, 17.1 Hz). m/z (ES⁺, 70V) 192 (M−NH₃).

[0233] Intermediate 21

[0234] Ethyl (R)-3-(4-Aminophenyl)-3-(tert-butoxycarbonylamino)propanoate

[0235] Intermediate 20 (550 mg, 1.96 mmol) was dissolved in dioxan (10ml) and treated with sodium bicarbonate (1 g), water (10 ml) anddi-tert-butyl dicarbonate (427 mg) and the mixture stirred for 16 h.Water was added and the product extracted into EtOAc (×2), washed withbrine, dried (Na₂SO₄), filtered and concentrated to give the crudeproduct which was purified by column chromatography (S)O₂; CH₂Cl₂/MeOH20:1) to give the title compound (296 mg, 49%) as an oil. δ_(H) (CDCl₃)7.07 (2H, d, J 8.3 Hz), 6.64 (2H, d, J 8.3 Hz), 5.28 (1H, br s), 4.99(1H, m), 4.05 (2H, q, J 7.1 Hz), 2.82 (1H, dd, J 5.1, 6.5 Hz), 2.73 (1H,dd, J 15.1, 6.5 Hz), 1.42 (9H, s), 1.17 (3H, t, J 7.1 Hz). m/z (ES⁺,70V) 331 (MNa⁺).

[0236] Intemediate 22

[0237] Ethyl(3R)-3-[(tert-Butoxycarbonylamino)]-3-[4-(2,6-naphthyridin-1-ylamino)phenylpropanoate

[0238] Intermediate 21 (250 mg, 0.81 mmol) in 2-ethoxyethanol (2 ml) wastreated with 1-chloro-2,6-naphthyridine (134 mg) and heated at 120° for15 mins, then 100° C. for 1 h, then cooled and concentrated. The residuewas extracted into EtOAc (x 3), washed with sodium bicarbonate solution,brine, dried (Na₂SO₄), filtered and concentrated to give the crudeproduct. The products were purified by column chromatography (S)O₂;CH₂Cl₂/MeOH 50:1-20:1-10:1) to give the deprotected compound (106 mg,30%) as a brown gum and the title compound (98 mg, 36%) as a yellow gum.δ_(H) (CDCl₃) 9.18 (1H, s), 8.66 (1H, d, J 5.9 Hz), 8.20 (1H, d, J 58Hz), 7.73 (1H, d, J 5.9 Hz), 7.65 (2H, d, J 8.5 Hz), 7.30 (2H, d, J 8.5Hz), 7.19 (1H, d, J 5.8 Hz), 5.47 (1H, m), 5.08 (1H, m), 4.09 (2H, q, J7.1 Hz), 2.83 (2H, t, 16.4 Hz), 1.44 (9H, s), 1.20 (3H, t, 17.1 Hz). m/z(ES⁺, 70V) 437 (MH⁺).

[0239] Intermediate 23

[0240] Ethyl (R)-3-Amino-3-[4-(2,6-naphthyridin-1-ylamino)phenylpropanoate

[0241] Intermediate 22 (100 mg, 1 mmol) was dissolved in EtOAc (5 ml)and saturated with HCl gas. The reaction mixture was stirred to give aprecipitate which was filtered and dried to give the title compoundwhich was combined with the material isolated from the previousreaction. 5H (CDCl₃) 9.18 (1H, s), 8.65 (1H, d, J 5.9 Hz), 7.65 (2H, d,J 8.5 Hz,), 7.37 (2H, d, J 8.5 Hz), 7.19 (1H, d, J 5.7 Hz), 4.44 (1H, t,J 6.8 Hz), 4.15 (2H, q, J 7.1 Hz), 2.68 (2H, d, J 6.8 Hz), 1.25 (3H, t,J 1-7.1 Hz).

[0242] Intermediate 24

[0243] N—BOC—O-(2-Pyrimidinyl)-L-tyrosine methyl ester

[0244] A solution of N—BOC-L-tyrosine methyl ester (3.0 g, 10.2 mmol) inDMF (5 ml) was added to a suspension of NaH (60% in oil, 11.2 mmol, 447mg) in DMF (10 ml). After 10 min, a solution of 2-chloropyrimidine (11.2mmol, 1.28 g) in DMF (3 ml) was added and the mxiture stirred overnight.The reaction was quenched with water, diluted EtOAc and washed withwater and brine. The EtOAc layer was dried (Na₂SO₄) and concentrated invacuo. Purification by column chromatography [SiO₂, EtOAc/hexane, 1:1]gave the title compound. δ_(H) (DMSO d6) 8.62 (2H, d, J 4.8 Hz), 7.37(1H, d, J 8.1 Hz), 7.28 (2H, d, J 8.4 Hz), 7.24 (1H, t, J 4.8 Hz), 7.09(2H, d, J 8.4 Hz), 4.18 (1H, m), 3.01 (1H, dd, J 13.8, 4.6 Hz), 1.33(9H, s).

[0245] Intermediate 25

[0246] O-(2-Pyrimidinyl)-L-tyrosine methyl ester hydrochloride

[0247] Removal of BOC group from Intermediate 24 (HCl/EtOAc) gave thetitle compound as a white solid. δH (DMSO d₆) 8.69 (3H, m), 8.63 (2H, d,J 4.9 Hz), 7.31-7.25 (3H, m), 7.15 (2H, d, J 8.6 Hz), 4.30 (1H, m), 3.69(3H, s), 3.19 (1H, dd, J 14.5, 6.4 Hz), 3.12 (1H, dd, J 14.3, 7.2 Hz).

[0248] Intermediate 26

[0249] N BOC-O-(3,5-Dichloroisonicotinoyl)-L-tyrosine methyl ester

[0250] A solution of N-BOC-L-tyrosine methyl ester (2.95 g, 10 mmol) inTHF (10 ml) was added to a suspension of NaH (60% in oil, 11 mmol, 440mg) in THF (30 ml) at 0°. After 10 min, a solution of3,5-dichloroisonicotinoyl chloride (11 mml, 2.32 g) in THF (10 ml) wasadded and the mixture stirred at RT for 4 h. NH₄Cl (aq) was added andthe mixture extracted with DCM. The DCM extracts were dried (Na₂SO₄) andconcentrated in vacua. Recrystallisation (EtOAc/hexane) gave the titlecompound as white crystals (3.61 g, 77%). δH (DMSO d₆) 8.89 (2H, s),7.39 (2H, d, J 8.5 Hz), 7.32 (1H, d, J 8.2 Hz), 7.23 (2H, d, J 8.5 Hz),4.21 (1H, m), 3.62 (3H, s), 3.05 (1H, dd, J 13.8, 4.9 Hz), 2.89 (1H, dd,J 13.8, 10.5 Hz), 1.31 (9H). m/z (ES⁺, 70V) 410 (M++Na).

[0251] Intermediate 27

[0252] O-(3,5-Dichloroisonicotinoyl)-L-tyrosine methyl esterhydrochloride

[0253] Intermediate 26 (3.61 g) in EtOAc (150 ml) was treated withHCl/EtOAc (3m, 50 ml). The white precipitate produced was filtered offand dried to give the title compound as a white solid (1.93 g). δH (DMSOd₆) 8.90 (2H, s), 8.74 (3H, br), 7.42 (2H, d, J 8.5 Hz), 7.28 (2H, d, J8.6 Hz), 4.31 (1H, m), 3.67 (3H, s), 3.25 (1H, dd, J 14.2, 6.0 Hz), 3.17(1H, dd, J 14.1, 7.2 Hz). m/z (ES⁺, 70V) 369 (MH⁺).

[0254] Intermediate 28

[0255]3-Butyl4-methoxy-3-cyclobutene-1,2-dione

[0256] n-BuLi (8.13 ml of a 1.6M solution in hexane, 13 mmol) was addedslowly to a solution of 3,4-dimethoxy-3-cyclobutene-1,2-dione (1.42 g,10 mmol) in THF (100 ml) at −78°. After 2 h, trifluoroacetic anhydride(2.12 ml, 15 mmol) was added. After a further 30 min the cold solutionwas poured into NH₄Cl(aq) (100 ml) and EtOAc (100 ml) and stirred well.The aqueous layer was extracted with EtOAc. The organic extracts werewashed with brine, dried (Na₂SO₄) and concentrated in vacuo. Columnchromatography (S)O₂, EtOAc/hexane, 30:70) gave the title compound as ayellow oil (803 mg, 48%). δH (CDCl₃) 4.42 (3H, s), 2.60 (2H, t, 17.6Hz), 1.71-1.61 (2H, m), 1.44-1.32 (2H, m), 0.94 (3H, t, J 7.3 Hz). m/z(ES⁺, 70V) 169 (MH⁺).

[0257] Intermediate 29

[0258] Methyl(Z)-2-[(tert-butoxycarbonyl)amino]-3-(3-methoxy-4-nitrophenyl)-2-propenoate

[0259] Activated manganese IV oxide (26 g) was added to a mixture of3-methoxy-4-nitrobenzylalcohol (5.26 g, 28.7 mmol),N-(t-Butyloxycarbonyl)<-(diethylphosphono)glycine methylester (describedin WO99/47547) (8.91 g, 27.4 mmol) and DBU (4.29 ml, 28.7 mmol) in DCM(150 ml) at 00. The mixture was stirred at RT overnight then filtered.The filtrate was washed with dil. HCl, dried (Na₂SO₄) and evaporated invacuo. Recrystallisation from MeOH gave the title compound as pale browncrystals (4.6 g). 6H (DMSO d₆) 8.94 (1H, br s), 7.91 (1H; d, J 18.4 Hz),7.56 (1H, d, J 1.5 Hz), 7.36 (1H, dd, 18.5, 1.3 Hz), 7.12 (1H, br s),3.92 (3H, s), 3.75 (3H, s), 1.37 (9H, s). M/Z (ES⁺, 70V) 375 (M⁺+Na).

[0260] Intermediate 30

[0261] Methyl3-(4-amino-3-methoxyphenyl)-2-[(tert-butoxycarbonyl)amino]-2-propanoate

[0262] A mixture of Intermediate 29 (2.30 g, 6.53 mmol) and palladium oncharcoal (10% Pd on carbon, 230 mg) in MeOH (65 ml) was stirred under ahydrogen atmosphere at RT overnight. The catalyst was filtered off andthe filtrate concentrated in vacuo. Recrystallisation (Et₂O/hexane) gavethe title compound as dark pink needles (1.62 g, 77%). δH (DMSO d₆) 7.12(1H, d, J 7.9 Hz), 6.65 (1H, s), 6.51 (2H, s), 4.52 (1H, s), 4.49 (1H,s), 4.07 (1H, m), 3.72 (3H, s), 3.59 (3H, s), 2.81 (1H, dd, J 13.7, 5.4Hz), 2.69 (1H, dd, J 13.1, 9.5 Hz), 1.32 (9H, s). m/z (ES⁺, 70V) 347(MNa⁺).

[0263] Intermediate 31

[0264] Methyl3-{4-[(6,7-dimethoxy-4-quinazolinyl)amino]-3-methoxyphenyl}-2-[(tert-butoxycarbonyl)amino]propanoate

[0265] A mixture of Intermediate 30 (486 mg, 15 mmol),4-chloro-6,7-dimethoxy quinazoline (337 mg, 1.5 mmol) anddiisopropylethylamine (2610, 1.5 mmol) in ethoxyethanol (1.5 ml) washeated at 120° for 24 h. The mixture was diluted with DCM, washed withdil. HCl and water, dried (Na₂SO₄) and concentrated in vacuo. Columnchromatography (S)O₂: MeOH/DCM, 5:95) gave the title compound as a browngum (720 mg, 94%) δH (DMSO d6) 9.10 (1H, s, ArNH), 8.34 (1H, d, J 1.0Hz), 7.85 (1H, d, J 1.4 Hz), 7.47-7.44 (2H, m), 7.40 (1H, d, J 8.0 Hz),7.47-7.44 (2H, m), 7.40 (1H, d, J 8.0 Hz), 7.20 (1H, s), 7.07 (1H, s),6.91 (1H, d, J 8.0 Hz), 4.344.28 (1H, m), 3.98 (3H, s), 3.98 (3H,s),3.82 (3H, s) 3.70 (3H, s), 3.09 (1H, dd, J 13.8, 5.0 Hz), 2.95 (1H,dd, J 13.7, 10.0 Hz), 1.42 (9H, s). m/z (ES⁺, 70V) 573 (MH⁺).

[0266] Intermediate 32

[0267] Methyl2-amino-3-{4-[(6,7-dimethoxy4-quinazolinyl)amino]-3-methoxyphenyl}propanoatehydrochloride Dry HCl was bubbled into a solution of Intermediate 31(715 mg, 1.4 mmol) in EtOAc (30 ml) for a few seconds. The mixture wasstirred at RT for 1 h. The precipitate was filtered off and dried togive the title compound as a brown solid (534 mg, 85%). δH (DMSO d₆,370K) 8.57 (1H, s), 8.23 (1H br s), 7.43 (1H, d, J 7.9 Hz), 7.15 (1H,s), 6.95 (1H, dd, J 8.0, 1.5 Hz), 4.28 (1H, dd, J 7.1, 6.2 Hz), 4.02(3H, s), 4.01 (2H, s), 3.82 (3H, s), 3.75 (3H, s), 3.31 (1H, dd, J 14.2,6.1 Hz), 3.24 (1H, dd, J 14.2, 7.1 Hz). m/z (ES⁺, 70V) 413 (MH⁺).

[0268] Intermediate 33

[0269] Methyl(S)-2-[(tert-butoxycarbonyl)amino]-3-{4-[2-(2,6-dichlorophenyl)ethynyl]phenyl}propanoate

[0270] Nitrogen was bubbled through a solution ofN—BOC-L-4-iodophenylalanine methyl ester (1.50 g, 3.69 mmol) in toluene(20 ml) and triethylamine (10 ml). Bis(triphenylphosphine)palladium (II)chloride (10 mol %, 260 mg) and copper (I) iodide (20 mol %, 140 mg)were added. A solution of 2,6-dichlorophenylacetylene (949 mg, 5.55mmol) in toluene (10 ml) was added by syringe-pump over 3 h. The mixturewas stirred at RT for a further 3 h. The mixture was diluted with EtOAc,washed with dil. HCl and brine, dried (Na₂SO₄) and evaporated in vacuo.Column chromatography (S)O₂; EtOAc/hexane, 20:80) gave the titlecompound as a brown gum (1.61 g, 97%). δH (DMSO d₆), 7.60-7.58 (2H, m),7.51 (2H, d, J 8.1 Hz), 7.42 (1H, dd, J 8.8, 7.4 Hz), 7.33 (2H, d, J 8.1Hz), 4.21 (1H, br m), 3.73 (3H, s), 3.04 (1H, dd, J 13.8, 5.0 Hz), 2.88(1H, dd, J 13.7, 10.0 Hz) and 1.31 (9H, s): m/z (ES⁺, 70V) 470 (M ⁺+Na).

[0271] Intermediate 34

[0272] Methyl(S)-2-amino-3-{4-[2-(2,6-dichlorophenyl)ethynyl]phenyl}propanoatehydrochloride

[0273] HCl gas was bubbled through a solution of the compound of Example33 (1.6 g, 3.57 mmol) in EtOAc (70 ml) for 5 min. The mixture wasstirred for 1 h at RT. The precipitate formed was filtered off andwashed with ether to give the title compound as an off-white solid (1.21g, 88%). δH (DMSO d6), 8.73 (3H, br s), 7.60 (2H, d, J 8.0 Hz), 7.56(2H, d, J 8.1 Hz), 7.44 (1H, dd, J 8.7, 7.6 Hz), 7.35 (2H, d, J 8.1 Hz),4.30 (1H, t, J 6.6 Hz), 3.68 (3H, s), 3.25 (1H, dd, J 14.2, 6.1 Hz),3.16 (1H, dd, J 14.0, 7.2 Hz); m/z (ES⁺, 70V) 348 (M ⁺+H).

[0274] Intermediate 35

[0275] 5-Methyl4-[3H]quinazolinone

[0276] 6-Methylanthranilic acid (5 g, 33 mmol) and formamidine acetate(0.4 g, 41 mmol) were refluxed in 2-ethyoxyethanol (50 ml) for 16 h. Oncooling the solvent was removed in vacuo, the residue slurried indiethyl ether, the solid filtered, washed with diethyl ether and driedto yield 3.6 g of the title compound. δH (DMSO d₆), 7.97 (1H, s), 7.60(1H, dd, J 7.9, 7.6 Hz), 7.43 (1H, d, J 8.0 Hz), 7.21 (1H, d, J 7.3 Hz),2.76 (3H, s); m/z (ES⁺ 70V)161 (MH⁺).

[0277] Intermediate 36

[0278] 4-Chloro-5-methylquinazoline

[0279] The compound of Intermediate 35 (4.1 g, 26 mmol) was refluxed inphosphorous oxychloride (60 ml) for 5 h. On cooling the phosphorousoxychloride was removed in vacuo and the residue quenched in ice coldsaturated sodium bicarbonate. The resulting mixture was extracted withEtOAc (3×50 ml), washed with brine, dried (Mg₂SO₄), the solvent removedand the residue purified by column chromatography (Silica 1:1ethylacetate/isohexane) to yield the title compound as white solid.δ_(H) (DMSO d₆), 8.5 (1H, s), 7.7 (1H, dd) 7.8 (1H, d), 7.3 (1H, m)

[0280] Intermediate 37

[0281]Ethyl-(S)-3-{4-([5-methyl-4-quinazolinyl]amino)phenyl}-2-(t-butoxy-carbonyl)aminopropanoate

[0282] Ethyl-(S)-3-(4-aminophenyl)-2-[(t-butoxycarbonyl)amino]propanoate(413 mg, 1.4 mmol) and Intermediate 36 (250 mg, 1.4 mmol) were heated atreflux in EtOH (10 ml). The solution was cooled, solvent removed invacuo, residue stirred in EtOAc (10 ml) and sat. sodium bicarbonate (10ml), organic layer isolated, washed with sodium bicarbonate, brine,dried (MgSO₄) and the solvent removed, to yield the title compound as anoff white solid (520 mg). δH (CDCl₃) 8.6 (1H, s), 7.8 (1H, br, s), 7.7(1H, d, J 7.8 Hz), 7.6 (2H, m), 7.3 (1H, d, J 7.2 Hz), 7.2 (2H, d, J 8.7Hz), 5.2 (1H, br m), 4.6 (1H, br m) 4.2 (2H, q, 7.2 Hz), 3.15 (2H, brm), 3.1 (3H, s), 1.4 (9H, s), 1.25 (3H, t, J 7.2 Hz).

[0283] Intermediate 38

[0284]Ethyl-(S)-3-(4-[(5-methyl4-quinazolinyl)amino]phenyl)-2-aminopropanoate

[0285] The compound of Intermediate 37 (1.1 g, 2.5 mmol) in DCM (4 ml)and trifluoroacetic acid (2 ml) was stirred for 1 h. The solution waspoured onto saturated sodium bicarbonate and extracted with EtOAc (×3).The extracts were washed with brine, dried (MgSO₄), solvent removed invacuo to give the title compound as yellow oil. δH (CDCl₃), 8.6 (1H, s),7.8 (1H, br s), 7.7 (1H, d, J 8.4 Hz), 7.6 (3H, m), 7.3 (3H, m) 4.2 (2H,q, J 7.2 Hz), 3.7 (1H, m), 3.1 (1H, dd, J 13.6, 8.4 Hz), 3.0 (1H, s),2.8 (1H, dd, J 13.6, 7.9 Hz), 1.3 (3H, t, J 7.2 Hz). m/z (ES⁺, 70V) 351(MH⁺)

[0286] Intermediate 39

[0287] Methyl-2-amino-5-(trifluoromethoxy)benzoate

[0288] A mixture of 2-Bromo-4-trifluoromethoxy aniline (2.7 g, 10.6mmol) palladium (II) acetate (360 mg,) triethylamine (9 ml) and 1,3-bis(diphenylphosphino) propane (651 mg) in anhydrous methanol (10 ml) andanhydrous dimethyl formamide (10 ml) were cooled in ice/methanol bath,and carbon monoxide gas was bubbled through for 10 min. The mixture washeated at 700 under a partially inflated balloon of carbon monoxide for17 h. On cooling nitrogen was bubbled through the solution to dispenseexcess carbon monoxide, and the mixture was poured onto water (50 ml)and EtOAc (50 ml), filtered through Celite®, the organic layer isolated,and aqueous phase was extracted with EtOAc. The organic layers werecombined, washed with water (×2), brine (×2), dried (MgSO₄), and thesolvent removed in vacuo. The residue was distilled and the fractionboiling at 170°, 0.08 mbar collected to yield 1.8 g of a yellow liquid.5H (CDCl₃), 7.7 (1H, m), 7.1 (1H, m), 6.6 (1H, d, 19.0 Hz), 3.9 (3H, s).

[0289] Intermediate 40

[0290] 6-(Trifluoromethoxy)-4-[3H]-quinazoline,

[0291] Prepared in a similar manner to the compound of intermediate 35from the compound of Intermediate 39. δ_(H) (DMSO d6), 8.1 (1H, s), 7.9(1H, s), 7.8 (2H, m).

[0292] Intermediate 41

[0293]4-Chloro-6-(trifluoromethoxy)quinazoline.

[0294] Prepared from the compound of Intermediate 40 in a similar mannerto that described for Intermediate 36. δ_(H) (CDCl₃), 9.1 (1H, s), 8.1(1H, d, J 9.2 Hz), 80 (1H, m), 7.8 (1H, m); m/z (EI⁺, 70V) 249/251.

[0295] Intermediate 42

[0296]Ethyl-(S)-3-(4-{[6-trifluoromethoxy4-quinazolinyl]amino}phenyl-2-[(t-butoxycarbonyl)amino]propanoate

[0297] Prepared from Intermediate 41 in a similar manner to thatdescribed for Intermediate 37. δ_(H) (CDCl₃), 8.7 (1H, s), 8.0 (1H, d, J9.1 Hz), 7.8 (1H, br s), 7.6 (314, m), 7.2 (2H, d, J 8.5 Hz), 5.0 (1H,br s) 4.5 (1H, br s), 4.2 (2H, q, J 7.2 Hz), 3.1 (2H, br s), 1.4 (9H,s), 1.2 (3H, t, J 7.2 Hz).

[0298] Intermediate 43

[0299] Ethyl(S)-3-(4-{[6-trifluoromethoxy)4-quinazolinyl]amino}phenyl)-2-aminopropanoate

[0300] Prepared from the compound of Intermediate 42 in a similar mannerto that described for Intermediate 38. δ_(H) (CDCl₃), 8.7 (1H, s), 7.9(1H, d, J 9.2 Hz), 7.7 (1H, br m), 7.6 (3H, m), 7.2 (2H, d, J 7.1 Hz),4.2 (2H, q, J 7.2 Hz), 3.8 (1H, m), 3.1 (1H, m), 2.9 (1H, m), 1.3 (3H,t, J 7.2 Hz); m/z (EI⁺, 70V) 421 (MH⁺)

[0301] Intermediate 44

[0302]3-Amino-4-methoxy-3-cyclobutene-1,2-dione

[0303] 3,4-Dimethoxy-3-cyclobutene-1,2-dione (1.3 g, 9.2 mmol) in ofMeOH (10.0 ml) was treated with aqueous ammonia (10.0 ml of a 2.0Msolution) and stirred at ambient temperature for 2 h. The yellowprecipitate thus formed was recovered by filtration, washed with MeOHand Et₂O and dried in vacuo to afford the title compound (0.87 g, 75%)as an amorphous yellow powder δH (d⁶ DMSO) 8.32 (2H, br s), 4.28 (3H,s). m/z (ES⁺, 70V) 127 (MH⁺).

[0304] Intermediate 45

[0305]Methyl-(S)-3-{4-[(2-chloro-6,7-dimethoxy-4-quinazolinyl)amino]phenyl}-2-[(t-butoxycarbonyl)amino]propanoate

[0306] Prepared in a similar manner to the compound of Intermediate 9from methyl-(S)-3-(4-aminophenyl)-2-(N-t-butoxycarbonylamino)propanoateand 2,4-dichloro6,7-dimethoxyquinazoline. δH (CD₃OD) 7.72 (1H, s), 7.69(2H, d, J 8.4 Hz), 7.25 (2H, d, J 8.4 Hz), 7.05 (1H, s), 4.34 (1H, m),4.15 (2H, m, J 7.1 Hz), 4.00 (3H, s), 3.96 (3H, s), 3.08 (1H, m), 2.97(1H, m), 1.40 (9H, s), 1.23 (3H, t, J 7.1 Hz). m/z (ESI⁺ 531 (MH⁺).

EXAMPLE 1

[0307] Ethyl(S)-3-[4-(3,5-dichloro-4-pyridylcarboxamido)phenyl]-242-isopropoxy-3,4-dioxocylobut-1-enylamino)propanoate

[0308] A solution of Intermediate 3 (2.1 g, 5 mmol) in EtOH (25 ml) wastreated with DIPEA (0.96 ml, 5.5 mmol) and3,4-diisopropoxy-3-cyclobutene-1,2-dione (1.1 g; 5.5 mmol) and heated toreflux for 16 h. The reaction mixture was cooled and concentrated invacuo. The residue was taken up in EtOAc (50 ml) and washed with 10%aqueous citric acid (2×50 ml), NaHCO₃ solution (2×30 ml) and brine (30ml), dried (MgSO₄) and the solvent evaporated in vacuo to give a paleyellow oil, which was purified by column chromatography(S)O₂,EtOAc:hexane 1:1) to give the title compound as a white foam 1.62g, 62%). δH (DMSO d⁶), 10.45 (1H, s), 8.69 (2H, s), 8.52 (1H, d, J 8.4Hz), 7.57 (2H, d, J 7.6 Hz), 7.25 (2H, d, J 7.6 Hz), 5.22 (1H, m), 4.69(1H, m), 4.19 (2H, q, J 7.1 Hz), 3.25 (1H, dd, J 14.3, 5.2 Hz), 3.07(1H, dd, J 14.3, 9.4 Hz), 1.38 (6H, dd, J 6.2, 3.9 Hz), 1.23 (3H, t, J7.1 Hz).

EXAMPLE 2

[0309]Ethyl-(S)-3-[4-(3,5-dichloro4-pyridylcarboxamido)phenyl]-2-(2-[3-methoxypropylamine]-3,4-dioxocyclobut-1-enylamino)propanoate

[0310] A solution of the compound of Example 1 (1.55 g, 2.99 mmol) inEtOH (25 ml) was treated with 3-methoxypropylamine (0.34 ml, 3.3 mmol)and stirred for 16 h at RT. The white solid was isolated by filtration,and washed with cold Et₂O (3×10 ml) to give the title compound (1.38 g,84%). δH (DMSO d⁶),10.89 (1H, s), 8.80 (2H, s), 7.59 (2H, d, J 8.4 Hz),7.25 (2H, br m), 7.18 (2H, d, J 18.4 Hz), 4.99 (1H, m), 4.18 (2H, q, J7.1 Hz), 3.54 (2H, m), 3.37 (2H, t, J 6.3 Hz), 3.23 (3H, s), 3.16.(1H,m), 3.06 (1H, m), 1.75 (2H, q. 16.3 Hz), 1.22 (3H, t, J 7.1 Hz). m/z(ES⁺, 70V) 549 (MH⁺).

EXAMPLE 3

[0311](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(3-methoxypropylamino]-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0312] A solution of the compound of Example 2 (1.30 g, 2.48 mmol) inTHF (40 ml) and water (25 ml) was treated with LiOH.H₂O (125 mg, 2.98mmol) and stirred for 3 h at RT. The reaction mixture was concentratedin vacuo, and acidified to pH 2 with 1M hydrochloric acid. The resultingsolid was isolated by filtration, washed with water and dried in vacuoto give the title compound (1.15 g, 85%). δH (DMSO d⁶), 10.89 (1H, s),8.79 (2H, s), 7.58 (3H, m), 7.19 (2H, d, J 8.1 Hz), 4.92 (1H, m), 3.54(2H, m), 3.23 (3H, s), 3.16 (1H, dd, J 13.9, 5.1 Hz), 3.05 (1H, dd, J13.9, 7.4 Hz) and 1.74 (2H, t, J 6.4 Hz). m/z (ES⁺, 70V) 521 (MH⁺).

EXAMPLE 4

[0313]Ethyl-(S)-3-[4-(3,5-dichloro-4-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0314] A solution of the compound of Example 1 (1 g, 1.93 mmol) in EtOH(25 ml) was treated with n-propylamine (0.18 ml, 2.12 mmol) and stirredat RT for 16 h. The resulting white solid was isolated by filtration andwashed with cold Et₂O (2×20 ml) to give the title compound (0.689, 68%).δH (DMSO d⁶), 10.87 (1H, s), 8.78 (2H, s), 7.57 (4H, m,), 7.16 (2H, d, J8.3 Hz), 4.97 (1H, m), 4.16 (2H, q, J 7.1 Hz), 3.44 (2H, m), 3.11 (2H,m), 1.50 (2H, m), 1.20 (3H, t, J 7.1 Hz), 0.86 (3H, t, J 7.1 Hz). m/z(ES⁺, 70V) 519 (MH⁺).

EXAMPLE 5

[0315] (S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-deoxocyclobut-1-enylamino)propanoicacid.

[0316] The title compound (0.67 g, 99%) was prepared from the compoundof Example 4 (0.66 g, 1.27 mmol) in a similar manner to the compound ofExample 3. δH (DMSO d⁶), 10.51 (1H, s), 8.71 (2H, s), 7.56 (2H, d, J 8.3Hz), 7.36 (1H, m), 7.31 (1H, d, J 9.0 Hz), 7.22 (2H, d, J 8.3 Hz), 4.96(1H, m), 3.49 (2H, q, 16.7 Hz), 3.20 (1H, dd, J 14.1, 5.6 Hz), 3.09 (1H,dd, J 14.1, 7.4 Hz), 1.57 (2H, m), 0.92 (3H, t, J 7.4 Hz). m/z (ES⁺,70V) 491 (MH⁺).

EXAMPLE 6

[0317] Ethyl(S)-3-[4-(3,5-dichloro4-pyridylcarboxamido)phenyl]-2-[(2-tert-bukyl)-3,4-dioxo-1-cyclobutenylamino]propanoate

[0318] A mixture of the compound of Intermediate 4 (392 mg, 2 mmol),Intermediate 3 (837 mg, 2 mmol) and DIPEA (348 μl, 2 mmol) in abs.ethanol (20 ml) was heated at reflux for 24 h. The solvent was removedin vacuo and the residue dissolved in DCM, washed with HCl (1M), dried(Na₂SO₄) and evaporated in vacuo. Column chromatography (S)O₂; MeOH/DCM,5:95) gave the title compound as a yellow foam (741 mg, 72%). δH (DMSOd₆), 10.83 (1H, s), 8.77 (2H, s), 8.54 (1H, d, J 8.6 Hz,), 7.54 (2H, d,J 8.4 Hz), 7.23 (2H, d, J 8.5 Hz), 5.01 (1H, m), 4.17 (2H, q, J 7.1 Hz),3.25 (1H, dd, J 4.6 Hz), 3.04 (1H, dd, J 13.7, 10.9 Hz), 1.21 (9H, s),1.21 (3H, t, J 7.1 Hz) m/z (ES⁺, 70V) 518 (M⁺+H).

EXAMPLE 7

[0319](S)-3-[4-[3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[(2-tert-butyl)-3,4-dioxo-1-cyclobutenylamino]propanoate

[0320] Lithium hydroxide monohydrate (66 mg, 1.56 mmol) was added to thecompound of Example 6 (735 mg, 1.42 mmol) in THF (14 ml) and water (14ml). After 2.5 h at RT the THF was removed in vacuo. The aqueous residuewas acidified (pH1, 1M HCl) and the precipitate filtered off, washedwith water and dried to give the title compound as a pale brown solid(625 mg, 90%). δH (DMSO d₆), 13.29 (1H, br s), 10.85 (1H, s), 8.78 (2H,s), 8.49 (1H, d, J 9.2 Hz), 7.55 (2H, d, J 8.5 Hz), 7.24 (2H, d, J 8.5Hz), 4.95 (1H, ddd, J 11.0, 9.3, 4.2 Hz), 3.28 (1H, dd, J 13.8, 4.1 Hz),3.04 (1H, dd, J 13.7, 1.1 Hz), 1.22 (9H, s); m/z (ES⁺, 70V) 490 (M ⁺+H).

EXAMPLE 8

[0321] Methyl(S)-3-{4-[(3,5-dichloroisonicotinoyl)oxy]phenyl}-2-(2-propylamino-3,4-dioxocyclbut-1-enylamino)propanoate

[0322] In a similar manner to that described for Example 1 and Example 2the title compound was prepared from the compound of Intermediate 27 asa white solid. δH (DMSO d₆, 390K) 8.81 (2H, s), 7.36 (2H, d, J 8.7 Hz),7.26 (2H, d, J 8.7 Hz), 5.11-5.05 (1H, m), 3.78 (3H, d), 3.52-3.47 (2H,m) 3.29 (1H, dd, J 14.2, 5.9 Hz), 3.18 (1H, dd, J 14.2, 9.7 Hz),1.63-1.54 (2H, m), 0.93 (3H, t, 17.4 Hz,). m/z (ES⁺, 70V) 506 (MH⁺).

EXAMPLE 9

[0323](S)-3-{(4-[(3,5-Dichloroisonicotinoyl)oxy]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0324] In a similar manner to that described for Example 3 the titlecompound was prepared from Example 8 as a white solid. δ_(H) (DMSO d₆,390K) 13.31 (1H, br), 8.80 (2H, s), 7.38 (2H, d, J 8.6 Hz), 7.25 (2H, d,J 8.6 Hz), 5.0-4.98 (1H, m), 3.52-3.47 (2H, m,) 3.29 (1H, dd, J 14.2,5.7 Hz), 3.16 (1H, dd, J 14.2, 7.5 Hz), 2.51-2.50 (2H, m), 0.93 (3H, t,J 7.4 Hz). m/z (ES⁺, 70V) 494 (MH⁺).

EXAMPLE 10

[0325] Ethyl(S)-3-[4-(3,5-dichloro-4-pyridylcarboxamido)phenyl]-2-(2-butyl-3,4-dioxo-1-cyclobutenylamino)propanoate

[0326] A mixture of Intermediate 28 (336 mg, 2 mmol), Intermediate 3(837 mg, 2 mmol) and DIPEA (700 μl, 4 mmol) in EtOH (2 ml) was heated atreflux for 2 h. The solvent was removed in vacuo. The residue wasdissolved in DCM (150 ml), washed with dil. HCl, dried (Na₂SO₄) andconcentrated in vacuo. Column chromatography (S)O₂: MeOH/DCM, 5:95) gavethe title compound as a yellow foam (904 mg, 87%). δH (DMSO d_(6, 390)K)10.39 (1H, br s), 8.68 (2H, s), 8.59 (1H, br d, J 7.8 Hz), 7.55 (2H, brs), 7.26 (2H, d, J 8.3 Hz), 4.84 (1H, br s), 4.21 (2H, q, J 7.1 Hz),3.28 (1H, dd, J 14.3, 5.3 Hz), 3.10 (1H, dd, J 14 3, 9.2 Hz), 2.5 (2H,m), 1.62-1.54 (2H, m), 1 38-1.29 92H), 1.24 (3H, t, J 7.1 Hz,CO₂CH₂CH₃), 0.91 (3H, t, J 7.3 Hz). m/z (ES⁺, 70V) 518 (MH⁺).

EXAMPLE 11

[0327] (S)-3-{4-(Dichloro-4-pyridylcarboxamido)phenyl]-2-(2butyl-3,4-dioxo-1-cyclobutenylamino)propanoic acid

[0328] In a similar manner to that described for Example 3 the titlecompound was prepared from the compound of Example 10 as a pale yellowsolid. 5H (DMSO d₆, 370K), 10.48 (1H, s), 8.70 (2H, s), 8.5 (1H, v br),7.55 (2H, d, J 7.8 Hz), 7.25 (2H, d, J 7.9 Hz), 4.85 (1H, v br),3.29-3.22 (1H, m), 3.09-3.03 (1H, m), 2.5 (2H, m), 1.57-1.51 (2H, m),1.36-1.27 (2H, m), 0.90 (3H, t, J 7.3 Hz). m/2 (ES, 70V) 490 (MH⁺).

EXAMPLE 12

[0329] Ethyl(S)-3-{4-[(2,6-naphthyridin-1-yl)amino]phenyl}2-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0330] A solution of Intermediate 7 (280 mg, 0.84 mmol) and3,4-diisopropoxy-3-cyclobuten-1,2-dione (200 mg, 1.01 mmol) in absoluteethanol (5 ml) was stirred at RT for 8 h then at 500 for 18 h. Thevolatiles were removed in vacuo and the residue chromatographed (Silica,80% EtOAc/Hexane to 100% EtOAc) affording the title compound as a dullyellow foam (250 mg, 63%). δH (CDCl₃) 9.18 (1H, s), 8.66 (1H, d, J 5.9Hz), 8.21 (1H, d, J 5.7 Hz), 7.72 (1H, d, J 5.9 Hz), 7.66 (2H, d, 18.5Hz), 7.22 (1H, obs. s), 7.20 (1H, d, 15.7 Hz), 7.14 (2H, d, J 8.5 Hz),6.37, 5.90, 5.18 and 4.60 (together 1H, br m's), 4.27 (2H, q, J 7.1 Hz),3.31-3.10 (2H, br m), 1.42 (3H, d, J. 6.2 Hz), 1.41 (3H, d, 16.2 Hz),1.32 (3H, t, J 7.1 Hz); m/z (ES⁺, 70V) 475 (MH⁺).

EXAMPLE 13

[0331] Ethyl(S)-3-{4-[(2,6-naphthyridin-1-yl)amino]phenyl}2-{[2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl]amino}propanoate

[0332] The compound of Example 12 (240 mg, 0.51 mmol) and diethylamine(74 mg, 105 μl, 1.01 mmol) in absolute ethanol (2 ml) was stirred at 45°under an atmosphere of N₂ for 18 h. The volatiles were removed in vacuoand the residue chromatographed (Silica, gradiant elution 1 to 3%EtOH/EtOAc) to afford the title compound as a yellow foam (240 mg, 97%).δH (CDCl₃) 9.17 (1H, s), 8.65 (1H, d, J 5.9 Hz), 8.19 (1H, d, J 5.7 Hz),7.78 (1H, d, J 5.9 Hz), 7.68 (2H, d, J 8.4 Hz), 7.48 (1H, s), 7.18 (1H,d, J 5.7 Hz), 7.13 (2H, d, J 8.4 Hz), 5.45-5.35 (2H, overlappingsignals), 4.25 (2H, q, J 7.1 Hz), 3.68-3.31 (4H, br m), 3.30-3.18 (2H,m), 1.31 (3H, t, J 7.1 Hz), 1.22 (6H, t, J 7.1 Hz); m/z (ES⁺, 70V) 488(MH⁺).

EXAMPLE 14

[0333](S)-3-{4-[(2,6-Naphthyridin-1-yl)amino]phenyl}2-{[2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl]amino}propanoicacid

[0334] The compound of Example 13 (230 mg, 0.47 mmol) was treated with asolution of LiOH.H₂O (25 ml, 0.60 mmol) in water (4 ml) and dioxan (4ml) at RT for 1.5 h. A few drops of AcOH were added and the volatilesremoved in vacuo. The residue was chromatographed [silica, gradiantelution, DCM (200 to 120), MeOH (20), AcOH (3), H₂O (2)] to afford theproduct as a yellow oil. Freeze-drying from aqueous MeOH afforded thetitle compound as a bright yellow amorphous solid (165 mg, 76%). δ_(H)(d₆ DMSO) 9.28 (1H, s), 9.20 (1H, s), 8.65 (1H, d, J 5.9 Hz), 8.37 (1H,d, J 5.8 Hz), 8.12 (1H, d, J 5.8 Hz), 7.78 (2H, d, J 8.5 Hz), 7.66 (1H,d, J 9.0 Hz), 7.26 (1H, d, J 5.8 Hz), 7.22 (2H, d, J 8.5 Hz), 5.15-5.05(1H, m), 3.70-3.30 (4H, br m), 3.22 (1H, dd, J 13.9, 4.0 Hz), 3.00 (1H,dd, J 13.9, 10.9 Hz), 1.09 (6H, t, J 7.1 Hz); m/z (ES⁺, 70V) 460 (MH⁺).

EXAMPLE 14A.

[0335](S)-3-{4-[(2,6-Naphthyridin-1-yl)amino]phenyl}-2-{[2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl]amino}propanoicacid, sodium salt

[0336] A solution of the compound of Example 14 (250 mg, 0.55 mmol) inwater (3 ml) and THF (2 ml) was treated with sodium hydroxide solution(0.1M, 5.5 mmol) and stirred for 10 mins. The solution was freeze driedto give the title compound as a bright orange solid (250 mg, 95%). δ_(H)(d₆ DMSO) 8.43 (1H, s), 8.06 (2H, s), 7.48 (1H, d, J 5.6 Hz), 7.16 (2H,d, J 8.3 Hz), 6.93 (2H, d, J 8.4 Hz), 5.88 (1H, d, J 5.6 Hz), 3.73 (1H,t, J 6.7 Hz), 3.88-3.83 (2H, m), 3.55-3.50 (2H, m), 2.86 (1H, dd, J13.3, 6.5 Hz), 2.67 (1H, m), 1.12 (6H, t, J 7.1 Hz). (ES⁺, 70V)460(MH⁺).

[0337] In a similar manner to that described for Examples 13 and 14 wereprepared the Examples 15 to 28:

EXAMPLE 15

[0338] Ethyl(S)-3-[4-(2,6-naphthyridin-1-ylamino)phenyl]2-[2-(piperadin-1-yl)-3,4-dioxocyclobut-1-enylamino]propanoate

[0339] δH (CDCl₃) 9.18 (1H, s), 8.67 (1H, d, J 5.9 Hz), 8.20 (1H, d, J5.9 Hz), 7.74 (1H, d, J 5.9 Hz), 7.67 (2H, d, J 8.5 Hz), 7.35 (1H, s),7.20 (1H, d, J 5.9 Hz), 7.13 (2H, d, J 8.5 Hz), 5.40 (2H, narrow m),4.25 (2H, q, J 7.2 Hz), 3.69-3.50 (4H, br m), 3.22 (2H, narrow m), 1.67(6H, narrow m), 1.31 (3H, t, J 7.2 Hz); m/z (ES⁺, 70V) (MH⁺) 500.

EXAMPLE 16

[0340](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[2-(piperidin-1-yl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0341] δH (d₆ DMSO) 9.29 (1H, s), 9.21 (1H, s), 8.65 (1H, d, J 5.9 Hz),8.38 (1H, d, J 5.9 Hz), 8.12 (1H, d, J 5.8 Hz), 7.77 (2H, d, J 8.4 Hz),7.76 (1H, obs. signal), 7.26 (1H, d, J 5.8 Hz), 7.21 (2H, d, J 8.4 Hz),5.07 (1H, narrow m), 3.72-3.48 (4H, br m), 3.20 (1H, dd, J 14.0, 4.1Hz), 2.98 (1H, dd, J 14.0, 10.6 Hz), 1.68-1.49 (6H, br m); m/z (ES⁺,70V) (MH⁺) 472.

EXAMPLE 17

[0342] Ethyl(S)-3-[4-(2,6-naphthyridin-1-ylamino)phenyl]-2-(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0343] δH (CDCl₃) 9.18 (1H, s), 8.70 (1H, d, 15.9 Hz), 8.15 (1H, s),7.85 (1H, br s), 7.64 (2H, d, J 8.3 Hz), 7.19-7.13 (3H, m), 5.40-5.30(1H, m), 4.35-4.20 (2H, m), 3.60-3.10 (6H, m), 1.65-1.55 (4H, m), 1.33(3H, t, J 7.1 Hz), 0.9 (6H, t, J 7.35 Hz), m/z (ES⁺, 70V) MH⁺ 516.

EXAMPLE 18

[0344](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0345] δH (d₆ DMSO, 370K) 9.19 (1H, s), 9.0 (1H, br s), 8.64 (1H, d,18.6 Hz), 8.34 (1H, d, J 5.9 Hz), 8.14 (1H, d, J 5.7 Hz), 7.79 (2H, d, J8.4 Hz), 7.25-7.21 (1H, m), 7.23 (2H, d, J 8.7 Hz), 7.05 (1H, br s),5.15 (1H, br s), 3.56-3.40 (4H, m), 3.27 (1H, dd, J 14.2, 4.9 Hz), 3.10(1H, dd, J 14.2, 9.4 Hz), 1.65-1.50 (4H, m), 0.86 (6H, t, 17.3 Hz), m/z(ES⁺, 70V) MH⁺ 488.

EXAMPLE 19

[0346](S)-3-[4-(2,6-naphthyridin-1-ylamino)phenyl]2-(2-tert-butyl-3,4-dioxocyclobut-1-enylamino)-propanoicacid

[0347] δ_(H) (d₆ DMSO) 9.29 (1H, s), 9.22 (1H, s), 8.67 (1H, d, J 5.8Hz), 8.51 (1H, d, J 9.1 Hz), 8.40 (1H, d, J 0.8 Hz), 8.38 (1H, d, J 0.8Hz), 8.13 (1H, dd, J 5.6, 1.3 Hz), 7.78 (2H, nr m), 7.26 (1H, d, 15.8Hz), 7.19 (1H, d, 18.6 Hz), 4.95 (1H, brs), 3,4-3.2 (1H, m), 3.04 (1H,dd, J 13.6, 11.1 Hz), 1.23 (9H, s). m/z (ES⁺, 70V) (MH⁺) 445.2.

EXAMPLE 20

[0348](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[2-N-methyl-N-butylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0349] δH (d6 DMSO, 390K) 9.19 (1H, s), 9.08 (1H, s), 8.65 (1H, d, 15.9Hz), 8.35 (1H, d, J 5.9 Hz), 8.35 (1H, d, J 5.9 Hz), 8.14 (1H, d, J 5.7Hz), 7.78 (2H, d, J 8.3 HZ), 7.25-7.20 (3H, m), 5.06 (1H, br s),3.58-3.42 (2H, m), 3.24 (1H, dd, J 14.1, 4.7 Hz), 3.16 (3H, s), 3.06(1H, dd, J 14.1, 9.5 Hz), 1.54-1.50 (2H, m), 1.27 (2H, dd, J 15.1, 7.4Hz), 0.87 (3H, t, J 7.31 HZ). M/Z ES⁺, 70V) 474 (MH⁺).

EXAMPLE 21

[0350](S)-3-[4-(2,6-Naphthyridin-1-yl-N-methylamino)phenyl]-2-[2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0351] δ_(H) (d₆ DMSO; 350K) 9.23 (1H, d, J 1.0 Hz), 8.36 (1H, d, J 5.6Hz), 8.22 (1H, d, J 6.0 Hz), 7.49 (1H, dd, J 5.7, 0.9 Hz), 7.30 (1H, brd, 18.0 Hz), 7.21 (2H, d, J 8.5 Hz), 7.05 (1H, d, J 6.0 Hz), 6.93 (2H,d, J 8.5 Hz), 5.12-5.09 (1H, narrow m), 3.66-3.45 (4H, m), 3.49 (3H, s),3.24 (1H, dd, J 14.0-4.5 Hz), 3.03 (1H, dd, J 14.0, 10.1 Hz), 1.10 (6H,t, J 7.1 Hz) m/z (ES⁺, 70V) 474 (MH⁺).

EXAMPLE 22

[0352](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[(2,5-dimethyl-3-pyrrolin-1-yl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0353] δH (DMSO d₆, 350K); 9.19 (1H, d, J 0.9 Hz), 9.09 (1H, s), 8.65(1H, d, J 5.9 Hz), 8.35 (1H, d, J 5.9 Hz), 8.14 (1H, d, J 5.7 Hz), 7.78(2H, d, J 8.3 Hz), 7.26-7.18 (4H, m), 5.90 (2H, s), 5.09 (1H, br s),4.85 (2H, q, J 12.8, 6.4 Hz), 3.27 (1H, dd, J 14.1, 4.8 Hz), 3.11 (1H,dd, J 14.1, 9.5 Hz), 1.35 (3H, d, J 6.4 Hz), 1.31 (3H, d, J 16.4 Hz),m/z (ES⁺, 70V) 484 (MH⁺).

EXAMPLE 23

[0354](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]2-[2-(N-methyl-N-propylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0355] δH (DMSO d₆, 350K), 9.20 (1H, s), 9.10 (1H, s), 8.65 (1H, d, J5.85 Hz), 8.35 (1H, d, J 5.92 Hz), 8.14 (1H, d, J 5.68 Hz), 7.79 (2H, d,J 8.03 Hz), 7.36 (1H, d, J 9.0 Hz), 7.26-7.22 (3H, m), 5.16 (1H, br s),3.50-3.39 (2H, m), 3.25 (1H, dd, J 14.09, 4.83 Hz), 3.17 (3H, s), 3.07(1H, dd, J 14.1, 9.9 Hz), 1.61-1.52 (2H, m), 0.84 (3H, t, J 7.35 Hz);m/z (ES⁺, 70V) 460.(MH⁺).

EXAMPLE 24

[0356](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[(2-(S)-(2-methoxymethyl)pyrrolidin-1-yl]-3,4-dioxocyclobut-1-enyl)amino[propanoicacid

[0357] δH (DMSO d_(6, 350)K) 9.20 (1H, s), 9.10 (1H, s), 8.65 (1H, d, J5.9 Hz), 8.35 (1H, d, J 5.9 Hz), 8.14 (1H, d, 0.15.7 Hz), 7.80 (2H, d, J8.3 Hz), 7.27-7.20 (4H, m), 5.07 (1H, br s), 4.20 (1H, d, J 5.2 Hz),3.85-3.64 (2H, m), 3.35-3.32 (2H, m), 3.25 (3H, s), 3.25-3.01 (2H, m),2.03-1.75 (4H, m); m/z (ES+, 70V), 502 (MH⁺).

EXAMPLE 25

[0358](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[2-(N-ethyl-N-iso-propylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0359] δH (DMSO d_(6, 350)K); 9.20 (1H, s), 9.09 (1H, s), 8.64 (1H, d, J5.9 Hz), 8.35 (1H, d, J 5.9 Hz), 8.15 (1H, d, J 5.7 Hz), 7.78 (2H, d, J8.3 Hz), 7.26-7.20 (4H, m), 5.18 (1H, brs), 4.44-4.37 (1H, m), 3.45 (2H,q, J 7.2, 2.4 Hz), 3.25 (1H, dd, J 14.1, 4.7 Hz), 3.08 (1H, dd, J 14.1,9.8 Hz), 1.20 (6H, q, J 6.7, 3.3 Hz), 1.14 (3H, t, 17.1 Hz), m/z (ES⁺,70V), 474 (MH⁺).

EXAMPLE 26

[0360](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[2-(methyl-N-iso-propylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0361] δH (DMSO d₆, 350K) 9.20 (1H, s), 9.09 (1H, s), 8.65 (1H, d, J 5.9Hz), 8.35 (1H, d, 15.9 Hz), 8.14 (1H, d, J 5.6 Hz), 7.79 (2H, d, J 8.3Hz), 7.38 (1H, d, J 8.1 Hz), 7.26-7.22 (3H, m), 5.12 (1H, br s),4.46-4.40 (1H, m), 3.25 (1H, dd, J 4.1, 4.8 Hz), 3.05 (1H, dd, J 14.2,4.6 Hz), 3.06 (3H, s). 1.82 (3H, d, J 2.58 Hz, 1.65 (3H, d, J 2.6 Hz);m/z (ES⁺, 70V) 460 (MH⁺,).

EXAMPLE 27

[0362](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[2-(2,5-dimethylpyrrolidin-1-yl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0363] δH (DMSO d₆, 350K) 9.20 (1H, d, J 0.9 Hz), 9.10 (1H, s), 8.65(1H, d, J 5.9 Hz), 8.35 (1H, d, J 5.9 Hz), 8.14 (1H, d, J 5.7 Hz), 7.79(2H, d, J 8.3 Hz), 7.26-7.23 (3H, m), 3.26 (1H, dd, J 14.2, 4.8 Hz), 3.1(1H, dd, J 14.2, 9.7 Hz), 2.15-2.09 (2H, m), 1.73-1.66 (2H, m), 1.28(3H, d, J 6.4 Hz), 1.25 (3H, d, J 6.4 Hz); m/z (ES⁺, 70V) 486 (MH⁺).

EXAMPLE 28

[0364](S)-3-[4-(2,6-Naphthyridin-1-ylamino)phenyl]-2-[2-(2-methylpiperdin-1-yl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0365] δH (DMSO d₆, 370K), 9.19 (1H, s), 9.03 (1H, s), 8.64 (1H, d, J5.8 Hz), 8.33 (1H, d, J 5.9 Hz), 8.14 (1H, d, J 5.6 Hz), 7.78 (2H, q, J8.4, 2.3 Hz), 7.25-7.22 (4H, m), 5.13 (1H, br s), 4.45 (1H, br s), 4.04(1H, d, J 13.7 Hz), 3.25-3.20 (2H, m), 3.11-3.05 (1H, m), 1.76-1.49 (6H,m), 1.24 (3H, q, J 6.9, 5.2 Hz), m/z (ES⁺, 70V) 486 (MH⁺).

EXAMPLE 29

[0366] Methyl(S)-3-[4-(2,6-naphthyrodin-1-yloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0367] To methyl(S)-2-(2-isopropoxy-3,4-dioxocyclobut-1-enylamino)-3-[4-(2,6-naphthyridin-1-yloxy)phenyl]-propanoate(prepared from the compound of Intermediate 8 in a similar manner to thecompound of Example 1) (0.20 g, 0.44 mmol) in methanol (3 ml) was added2 equivalents of diethylamine (0.09 ml) and the solution was stirred at65° overnight. The solution was cooled and then evaporated. The solidwas chromatographed (Silica, EtOAc/isohexane 50-100%) to afford thetitle com pound (0.15 g, 73%) as a white solid. δH (CDCl₃) 9.30 (1H, s),8.77 (1H, d, J 5.7 Hz), 8.19 (1H, d, 15.8 Hz), 8.08 (1H, d, J 5.79 Hz),7.44 (1H, d, J 5.8 Hz), 7.24-7.18 (4H, m), 5.46 (1H, m), 5.35 (1H, m),3.83 (3H, s), 3.70-3.40 (4H, br s), 3.31 (2H, d, J 5.3 Hz), 1.24 (6H, t,J 7.2 Hz). m/z (ES⁺, 70V) MH⁺ 475.

EXAMPLE 30

[0368](S)-3-[4-(2,6-Naphthyridin-1-yloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0369] The compound of Example 29 (0.137 g, 0.29 mmol) in dioxan (2 ml)and water (2 ml) was treated with LiOH.H₂O (0.02 g) and stirred at RTfor 4 h, a few drops of glacial acetic acid were added and the solutionwas then evaporated in vacuo. The product was chromatographed (Silica;DCM 200:MeH 20:HOAc 3:H₂O 2) to afford the title compound as anoff-white solid (0.10 g, 78%). δ_(H) (d₆ DMSO, 350K), 9.40 (1H, s), 8.76(1H, d, J 5.7 Hz), 8.15-8.09 (2H, m), 7.65 (1H, dd, J 5.8, 0.9 Hz), 7.37(1H, s), 7.36 (2H, d, J 8.6 Hz), 7.20 (2H, d, J 8.6 Hz), 5.15 (1H, brs), 3.59-3.51 (4H, m), 3.32 (1H, dd, J 4.1, 4.8 Hz), 3.13 (1H, dd, J14.1, 9.9 Hz), 1.14 (6H, t, J 7.1 Hz). m/z (ES⁺, 70V) MH⁺ 461.

[0370] The compounds of Examples 31 to 33 were prepared in a similarmanner to the compounds of Examples 29 and 30.

EXAMPLE 31

[0371](S)-3-[4-(2,6-Naphthyridin-1-yloxy)phenyl]-2-[2-piperidin-1-yl-3,4-dioxocyclobut-1-enylamino]propanoiacacid

[0372] δH (d₆ DMSO, 370K) 9.39 (1H, s), 8.76 (1H, d, J 5.7 Hz), 8.13(2H, nr m), 7.65 (1H, dd, J 5.7, 0.9 Hz), 7.34 (2H, d, J 8.6 Hz), 7.21(2H, d, J 8.6 Hz), 5.16 (1H, br s), 3.64-3.59 (5H, m), 3.31 (1H, dd, J14.1, 5.0 Hz), 3.12 (1H, dd, J 14.1, 9.6 Hz), 1.63-1.57 (5H, m); m/z(ES⁺, 70V) MH⁺ 473.

EXAMPLE 32

[0373]Methyl-(S)-3-[4-(2,6-naphthyridin-1-yloxy)phenyl]-2-(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0374] δH (d₆ DMSO) 9.41 (1H, s), 8.76 (1H, d, J 5.7 Hz), 8.14 (1H, d, J5.7 Hz), 8.07 (1H, d, J 5.7 Hz), 7.74 (1H, d, J 8.9 Hz), 7.67 (1H, d, J5.8 Hz), 7.33 (2H, d, J 8.5 Hz), 7.18 (2H, d, J 8.5 Hz), 5.23 (1H, m),3.72 (3H, s), 3.37 (5H, br m), 3.11 (1H, m), 1.48 (4H, br m), 0.80 (6H,t, J 7.3 Hz).

EXAMPLE 33

[0375](S)-3-[4-(2,6-Naphthyrodin-1-yloxy)phenyl]-2-(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1enylamino)propanoic acid

[0376] δH (d₆ DMSO 350K) 9.41 (1, d, J 1.0 Hz), 8.77 (1H d, J 8.7 Hz),8.14 (1H, d, J 5.7 Hz), 8.11 (1H, d, J 5.7 Hz), 7.67 (1H, dd, J 5.8, 0.9Hz), 7.35 (2H, d, J 8.6 Hz), 7.27 (1H, d, J 8.9 Hz), 7.21 (2H, d, J 8.6Hz), 5.20 (1H, m), 3.47 (4H, m), 3.33 (1H, dd, J 14.1, 4.8 Hz), 3.13(1H, dd, J 14.1, 10.0 Hz), 1.55 (4H, m), 0.86 (6H, t, J 7.4 Hz). m/z(ES⁺, 70V) 489 (MH⁺).

EXAMPLE 34

[0377]Methyl-(S)-3-{4-[2-(2,6-dichlorophenyl)ethynyl]phenyl}-2-[(2-isopropoxy-3,4-dioxo-1-cyclobutenyl)amino]propanoate

[0378] A mixture of the compound of Intermediate 34 (1.17 g, 3.04 mmol),3,4-diisopropoxy-3-cyclobutene-1,2-diene (632 mg, 3.19 mmol) and DIPEA(540 μl, 3.1 mmol) in MeOH (30 ml) was stirred at RT for 3 days. Thesolvent was removed in vacuo. The residue was dissolved in DCM, washedwith dil. HCl, dried (Na₂SO₄) and evaporated in vacuo. Columnchromatography (S)O₂; MeOH/DCM, 3:97) gave the title compound as ayellow gum (1.45 g, 98%). δH (DMSO d₆, 390K), 8.47 (1H, d, J 7.9 Hz),7.53-7.50 (3H, m), 7.38 (1H, dd, J 8.7, 7.5 Hz), 7.33 (2H, d, J 8.2 Hz),5.21 (1H, sept. J 6.2 Hz), 4.78-4.72 (1H, m), 3.72 (3H, s), 3.31 (1H,dd, J 14.2, 5.2 Hz), 3.13 (1H, dd, J 14.2, 9.4 Hz), 1.38 (3H, d, J 6.1Hz). 1.37 (3H, d, J 6.2 Hz); m/z (ES⁺, 70V) 486 (M ⁺+H).

EXAMPLE 35

[0379] Methyl(S)-3-{4-[2-(2,6-dichlorophenyl)ethynyl]phenyl}-2-[2-propylamino-3,4-dioxocyclobut-1-enylamino]propanoate

[0380] Propylamine (96 μl, 1.18 mmol) was added to a solution of thecompound of Example 34 (475 mg, 0.98 mmol) in MeOH (10 ml). The reactionmixture was stirred at RT overnight. Volatiles were removed in vacuo andthe resulting solid triturated with boiling MeOH. The solid was filteredoff to give the title compound as a white solid (335 mg, 71%). δH(DMSO-d₆, 390K) 7.57-7.53 (4H, m), 7.44-7.40 (1H, m), 7.33 (2H, d, J 8.3Hz), 7.3 (1H, br m) 7.2 (1H, br m), 5.10 (1H, m). 3.76 (3H, s),3.54-3.49 (2H, m), 3.30 (1H, dd, J 14.1, 5.9 Hz), 3.18 (1H, dd, J 14.1,7.7 Hz), 1.60 (2H, sept. J 7.1 Hz), 0.95 (3H, t, 17.4 Hz); m/z (ES⁺,70V) 485 (M ⁺+H).

EXAMPLE 36

[0381](S)-3-{4-[2-(2,6-Dichloropheny)ethynyl]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0382] Lithium hydroxide monohydrate (34 mg, 0.81 mmol) was added to thecompound of Example 35 (325 mg, 0.671 mmol) in a mixture of THF (7 ml)and water (7 ml). After 1 h at RT the THF was removed in vacuo. Theaqueous residue was acidified (pH 1-2, dil. HCl) and the precipitatedfiltered off, washed with water and dried to give the title compound asa yellow solid (315 mg, 90%). δH (DMSO d₆, 390K), 7.37-7.31 (4H, m),7.21 (1H, dd, J 8.6, 7.5 Hz), 7.14 (2H, d, J 8.4 Hz), 7.1 (2H, br m),4.82 (1H, m), 3.33-3.29 (2H, m), 3.11 (1H, dd, J 4.1, 5.7 Hz), 2.98 (1H,dd, J 14.1, 7.6 Hz), 1.39 (2H, sept. J 7.1 Hz). 0.75 (3H, t, J 7.4 Hz);m/z (ES⁺, 70V) 471 (M ⁺+H).

EXAMPLE 37

[0383] Methyl(S)-3-{4-[2-(2,6-dichlorophenyl)ethynyl]phenyl}-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0384] A mixture of the compound of Example 34 (470 mg, 0.969 mmol) anddiethylamine (401 μl, 3.88 mmol) in MeOH (10 ml) was heated at 50° C.overnight. The solvent was removed in vacuo and the residue purified bycolumn chromatography (S)O₂; MeOH/DCM, 5:95) to give the title compoundas a light brown foam (450 mg, 93%). δH (DMSO d₆, 390K), 7.55-7.50(4H,m), 7.42-7.35 (3H, m), 7.16 (1H, d, J 8.5 Hz), 5.63 (1H, m), 3.74 (3H,s), 3.55 (4H, q, J 7.1 Hz), 3.34 (1H, dd, J 14.2, 5.3 Hz), 3.20 (1H, dd,J 14.2, 9.4 Hz), 1.17 (6H, t, J 7.1 Hz); m/z (ES⁺, 70V) 499 (M ⁺+H).

EXAMPLE 38

[0385](S)-3-{4-[2-(2,6-Dichlorophenyl)ethynyl]phenyl}-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0386] Obtained as an off-white solid from the compound of Example 37 byester hydrolysis using the method described above for Example 36. δH(DMSO-d₆, 390K), 7.42-7.37 (4H, m), 7.29-7.24 (3H, m,), 6.91 (1H, br d,J 8.7 Hz), 3.43 (4H, q, J 7.1 Hz), 3.22 (1H, dd, J 14.2, 5.1 Hz), 3.06(1H, dd. J 14.2, 9.4 Hz), 1.04 (6H, t J 7.1 Hz). m/z (ES⁺, 70V) 485 (M⁺+H).

[0387] The compounds of Examples 39 to 44 were prepared frommethyl-(S)-3-(4-aminophenyl)-2-(N-t-butyloxycarbonylamino)propanoate andthe appropriate reagent in a similar manner to that described forIntermediate 3 then derivatised in a manner analogous to that describedfor Examples 1, 2 and 3.

EXAMPLE 39

[0388](S)-3-[4-(Benzylcarboxamido)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid δH (d₆ DMSO 390K); 9.25 (1H, s), 7.86 (1H, s), 7.44 (2H, d, J 8.4Hz), 7.40-7.15 (5H, m), 7.08 (2H, d, J 8.4 Hz), 7.0 (1H, d, 18.0 Hz),4.94 (1H, br s), 3.62 (2H, s), 3.47 (2H, nr m), 3.14 (1H, dd, J 14.1,5.7 Hz), 3.04 (1H, dd, J 14.1, 6.8 Hz), 1.57 (2H, dd, J 14.3, 7.1 Hz),0.92 (3H, t, J 7.3 Hz); m/z (ES⁺, 70V) 436 (MH⁺).

EXAMPLE 40

[0389](S)-3-[4-(2,4,6-Trifluorobenzylamino)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylaminopropanoicacid

[0390] δH (d₆ DMSO 390K) 7.88 (1H, s), 7.12 (1H, br s), 6.97 (1H, br s),6.92 (2H, d, J 8.3 Hz), 6.78 (2H, nr m), 6.58 (2H, d, J 8.3 Hz), 4.89(1H, br s)m 4.27 (2H, s), 3.46-3.48 (2H, nr m), 3.04 (1H, dd, J 14.18,5.7 Hz), 2.95 (1H, dd, J 14.2, 6.68 Hz), 1.62-1.53 (2H, Nr m), 0.92 (3H,t, J 7.38 Hz); m/z (ES⁺, 70V), 462 (MH⁺).

EXAMPLE 41

[0391](S)-3-[4(2,6-Dichlorobenzylamino)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1enylamino)propanoic acid

[0392] δH (d₆ DMSO) 7.26 (2H, s), 7.17 (1H, d, J 7.3 Hz), 7.14 (1H, brs), 6.95 (1H, br s), 6.8 (2H, d, J 8.4 Hz), 6.5 (2H, d, J 8.47 Hz), 4.70(1H, br s), 4.31 (3H, s), 3.13 (2H, m), 2.89 (1H, dd, J 14.2, 5.6 Hz),2.79 (1H, dd, J 14.2, 7.1 Hz), 2.85 (1H, br s), 1.44 (2H, dd, J 14.2,7.1 Hz), 0.76 (3H, t, J 7.4 Hz); m/z (ES⁺, 70V) 476 (MH⁺).

EXAMPLE 42

[0393] (S)-3-[4-(2,4,6-Trichlorobenzylamino)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoic acid

[0394] δ_(H) (d₆ DMSO) 7.55 (2H, s),7.23 (1H, br s), 7.09 (1H, br d, J8.4 Hz), 6.96 (2H, d, J 8.4 Hz), 6.66 (2H, d, J 8.5 Hz), 4.90 (1H, brs), 4.44 (2H, s), 3.48 (2H, m), 3.07 (1H, dd, J 14.1, 5.5 Hz), 2.95 (1H,dd, J 14.2, 7.2 Hz), 1.60 (2H, dd, J 14.3, 7.0 Hz), 0.93 (3H, t, J 7.4Hz); m/z (ES⁺, 70V) 509 (MH⁺).

EXAMPLE 43

[0395](S)-3-[4-(3-Chlorothiophen-2-ylcarboxamido)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0396] δH (d₆ DMSO) 9.50 (1H, s), 7.79 (1H, d, J 5.2 Hz), 7.57 (2H, d, J8.4 Hz), 7.21 (2H, d, J 8.4 Hz), 7.12 (1H, br s), 7.11 (1H, d, J 5.2Hz), 4.96 (1H, br s), 3.49 (2H, m), 3.25-3.02 (2H, m), 1.59 (2H, dd, J14.3, 7.1 Hz), 0.93 (3H, t, J 7.4 HZ), m/z (ES⁺, 70V) 461 (MH⁺).

EXAMPLE 44

[0397](S)-3-[4-(3-Chlorobenzo[b]thiophen-2-ylcarboxamido)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid δH (d6 DMSO, 400K) 9.98 (1H, s), 8.07 (1H, nr, m), 7.94 (1H, nr m),7.6 (5H, m), 7.23 (2H, d, 18.4 Hz), 7.1 (1H, br s), 4.98 (1H, br s), 3.5(2H, m), 2.35 (1H, dd, J 14.2, 5.7 Hz), 3.3 (1H, dd, J 14.2, 5.7 Hz),1.59 (2H, hex, J 7.3 Hz), 0.94 (3H, t, J 7.3 Hz). m/z (ES⁺, 70V) 512(MH⁺).

[0398] The compounds of Examples 45 to 47 were prepared from methyl(S)-3-(4-aminophenyl)-2-(N-t-butoxycarbonylamino)propanoate and theappropriate reagent in a similar manner to that described forIntermediate 6 then derivatised in a similar manner to that describedfor Examples 11, 13 and 14.

EXAMPLE 45

[0399](S)-3-[4-(Pyrimidin-2-ylamino)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0400] δH ((d₆ DMSO, 390K) 8.86 (1H, br s), 8.41 (2H, d, J 4.8 Hz), 7.64and 7.62 (2H, dd, J 1.8, 1.4 Hz), 7.15 (1H, br s), 7.12 (2H, d, J 8.6Hz), 6.77 (1H, t, J 4.8 Hz), 4.93 (1H, br s), 3.48 (2H, t, J 6.8 Hz),3.18 (1H, dd, J 14.1, 5.5 Hz), 3.05 (1H, dd, J 14.2, 7.3 Hz), 1.58 (2H,dd, J 14.2, 7.0 Hz), 0.92 (3H, t, 17.4 Hz), m/z (ES⁺, 70V) 396 (MH⁺).

EXAMPLE 46

[0401](S)-3-{4-[(2-Benzyl-6-chloropyrimidin4-yl)amino]phenyl}-2-{[2-N,N-diethylamino-3,4-dioxocyrclobut1enyl]amino}propanoic acid

[0402] δH (DMSO, 370K) 9.40 (1H, s), 7.48 (2H, d J 2.3 Hz), 7.38 (4H,s), 7.35-7.25 (2H, m), 7.24 (2H, d, 18.5 Hz), 6.64 (1H, s), 5.15 (1H, brs), 4.07 (2H, s), 3.60 (2H, q, J 7.2, 4.7 Hz), 3.3 0(1H, dd, J 14.2, 4.9Hz), 3.10 (1H, dd, J 14.1, 9.4 Hz), 1.2 (6H, t, J 7.1 Hz); m/z (ES⁺,70V) 534 (MH⁺).

EXAMPLE 47

[0403](S)-3-[4-(Quinolin4-ylamino)phenyl]-2-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0404] δH (DMSO, 390K) 8.48 (1H, d, J 5.2 Hz), 8.39 (1H, d, J 7.1 Hz),7.93 (1H, dd, J 8.4, 0.8 Hz), 7.71 (1H, d, J 5.4 Hz), 7.54-7.50 (1H, m),7.32 (2H, d, J 8.4 Hz), 7.24 (2H, d, J 8.5 Hz), 6.83 (1H, d, J 5.2 Hz),4.68 (1H, m), 3.70-3.50 (4H, m), 3.32 and 3.29 (1H, dd, J 13.8, 5.5 Hz),3.24 and 3.21 (1H, dd, J 13.8, 6.3 Hz), 1.23 (6H, t, J 7.2 Hz), m/z(ES⁺, 70V), 459 (MH⁺).

[0405] The compounds of Examples 48 to 55 were prepared fromN-BOC-L-tyrosine methyl ester and the appropriate reagent in the mannerdescribed for Intermediate 24 then derivatised in a manner analogous tothat described for Examples 12 to 14.

EXAMPLE 48

[0406] Methyl(S)-3-[4-(2,6-Dichlorobenzyloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0407] δH (DMSO d₆), 7.71 (1H, d, 19.0 Hz), 7.55-7.52 (2H, m), 7.47-7.42(1H, d), 7.18 (2H, d, J 8.7 Hz), 6.95 (2H, d, J 8.7 Hz), 5.17 (2H, s),5.15 (1H, m), 3.70 (3H, s), 3.55 (4H, br), 3.20 (1H, dd, J 4.6 Hz),3.01-2.93 (1H, m), 1.07 (6H, t, J 7.1 Hz); m/z (ES⁺, 70V) 505 (MH⁺).

EXAMPLE 49

[0408](S)-3-[4-(2,6-Dichlorobenzyloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0409] δH (DMSO d₆) 13.08 (1H, br), 8.31-8.24 (2H, m), 8.22-8.04 (1H,m), 8.02 (2H, d, J 8.8 Hz), 7.77 (2H, d, J 8.7 Hz), 6.07 (2H, s), 7.70(1H, br), 5.95 (1H, br m), 4.49-4.40 (4H, m), 4.05 (1H, dd, J 14.3, 5.1Hz), 3.89 (1H, dd, J 14.2, 9.1 Hz), 1.97 (6H, t, J 7.1 Hz); m/z (ES⁺,70V) 491 (MH⁺).

EXAMPLE 50

[0410] Methyl(S)-3-[4-(2,6-dichlorobenzyloxy)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0411] δH (DMSO d6) 7.60 (1H, br), 7.56 (2H, m), 7.47-7.42 (1H, m,),7.09 (2H, d, J 8.3 Hz), 6.97 (2H, d, J 8.7 Hz), 5.17 (2H, s), 4.99 (1H,m), 3.70 (3H, s), 3.70 (2H, m), 3.12 (1H, dd, J 5.2 partly obscured),1.54-1.47 (2H, m), 0.86 (3H, t, J 7.4 Hz). M/Z. (ES, 70V) 491 (MH⁺).

EXAMPLE 51

[0412](S)-3-[4-(2,6-Dichlorobenzyloxy)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0413] δH (DMSO d₆, 390K), 7.42-7.40 (2H, m), 7.35-7.31 (1H, m), 7.09(1H, br), 7.08-7.06 (2H, m), 6.89-6.86 (2H, m),5.17 (2H, s), 4.82 (1H,br), 3.39-3.38 (2H, m), 3.09 (1H, dd, J 14.2, 5.6 Hz), 2.96 (1H, dd, J14.2, 7.4 Hz), 1.52-1.47 (2H, m), 0.84 (3H, t, 17.4H₃); m/z (ES⁺, 70V)477 (MH⁺).

EXAMPLE 52

[0414] Methyl(S)-3-[4-12-pyrimidinyloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0415] δH (DMSO d₆, 390K) 860 (2H, d, J 4.8 Hz), 7.31 (2H, d, J 8.6Hz,), 7.20 (1 Ha, t, J 4.8 Hz). 7.10 (2H, d, J 8.6 Hz), 5.28-5.23 (1H,m), 3.74 (3H, s), 3.56 (4H, q, J 7.1 Hz), 3.31 (1H, dd, J 14.3, 5.4 Hz),3.17 (1H, dd, J 14.2, 9.2 Hz), 1.17 (6H, t, J 17.1 Hz); m/z (ES⁺, 70V)425 (MH⁺).

EXAMPLE 53

[0416](S)-3-[4-(2-Pyrimidinyloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0417] δH (DMSO d6, 390K) 13.10 (1H, br), 8.60 (2H, d, 4.8 Hz), 7.31(2H, d, J 8.7 Hz), 7.20 (1H, d, 14.8 Hz), 7.09 (2H, d, J 8.7 Hz), 6.97(1H, br), 5.18-5.17 (1H, m), 3.60-3.59 (4H, m), 3.31 (1H, dd, J 14.3,5.2 Hz), 3.16 (1H, dd, J 14.3, 9.1 Hz); m/z (ES⁺, 70V), 411 (MH⁺).

EXAMPLE 54

[0418] Methyl(S)-3-[4-(2-pyrimidinyloxy)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0419] δH (DMSO d₆, 390K) 8.61 (2H, d, J 4.8 Hz), 7.70 (1H, br), 7.55(1H, br), 7.26-7.19 (3H, m), 7 10 (2H, d, J 8.5 Hz), 5.02 (1H, m), 3.71(3H, s), 3.44 (2H, br), 3.18 (1H, dd, J 14.0, 5.4 Hz, CH _(A)H_(B)Ar),3.08 (1H, dd, J 14.0, 8.0 Hz, CH_(A) H _(B)Ar), 1.54-1.46 (2H, m,NHCH₂CH ₂CH₃), 0.86 (3H, t, J 7.4, NCH₂CH₂CH ₃); m/z (ES⁺, 70V) 411(MH⁺).

EXAMPLE 55

[0420](s)-3-[4-(2-Pyrimidinyloxy)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0421] δH (DMSO d₆, 390K) 8.67 (2H, d, J 4.8 Hz), 7.33 (2H, d, J 8.6Hz), 7.27 (1H, d, J 4.7 Hz), 7.16 (2H, d, J 8.6 Hz), 5.06-5.02 (1H, m),3.58-3.53 (2H, m), 3.31 (1H, dd, J 14.3, 5.6 Hz), 3.18 (1H, dd, J 14.2,7.5 Hz,), 1.67-1.62 (2H, m), 0.99 (3H, t, J 7.4 Hz); m/z (ES⁺, 70V) 397(MH⁺).

EXAMPLE 56

[0422] Methyl (S)-3-{4-[(3-phenyl-1quinazolinyl)amino]phenyl}-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0423] Intermediate 18 (518 mg, 1.3 mmol) was dissolved in MeOH (5 ml)and DIPEA base (0.5 ml), treated with3,4-diisopropoxy-3-cyclobutene-1,2-dione (309 mg) and stirred at RT for16 h. The solution was concentrated, dissolved in DCM (20 ml), washedwith water, dried (Na₂SO₄), filtered and concentrated. The crude productwas purified by column chromatography (S)O₂:CH₂Cl₂/MeOH 50:1) to givethe title compound (550 mg, 1.0 mmol, 79%) as a brown foamy solid. δH(DMSO) 8.44 (1H, m), 8.43 (2H, m), 7.80 (2H, m), 7.75 (2H, m), 7.50 (2H,m), 7.49 (2H, m), 7.33 (2H, d, J 8.6 Hz), 5.23 (1H, septet, J 6.2 Hz),4.80 (1H, m), 3.76 (3H, s), 3.30 (1H, dd, J 14.2, 5.3 Hz), 3.13 (1H, dd,J 14.2, 9.3 Hz), 1.38 (3H, d, 16.2 Hz), 1.37 (3H, d, 16.2 Hz); m/z (ESI,70V) 537 (MH⁺).

EXAMPLE 57

[0424] Methyl(S)-3-{4-[(2-phenyl-4-quinazolinyl)amino]phenyl}-[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl)amino]propanoate

[0425] The compound of Example 56 (550 mg, 1.0 mmol) and diethylamine(0.21 ml) in MeOH (5 ml) was stirred at RT for 16 h and the solutionthen concentrated. The residue was purified by column chromatography(S)O₂; DCM/MeOH 100:1) to give the title compound (375 mg, 0.68 mmol,68%) as a brown foamy solid. δH (DMSO, 390K) 8.45 (3H, m), 7.85 (4H, m),7.56 (1H, m), 7.48 (3H, m), 7.35 (2H, d, J 8.7H, 5.33 (1H, m), 3.76 (3H,s), 3.56 (2H, q, J 7.2 Hz), 3.54 (2H, q, J 7.2 Hz), 3.33 (1H, dd, J14.2, 5.3 Hz), 3.20 (1H, dd, J 14.2, 9.2 Hz), 1.17 (6H, t, J 7.1 Hz);m/z; (ES⁺, 70V) 550 (MH⁺).

EXAMPLE 58

[0426](S)-3-{-[2-Phenyl-4-quinazolinyl)amino]phenyl}-[2-N,N-diethylamino-3,4-dioxo-t-cyclobutenyl)amino]propanoicacid

[0427] Example 57 (360 mg, 0.66 mol) was dissolved in THF (2 ml) andwater (2 ml) and treated with lithium hydroxide (41 mg). The solutionwas stirred at RT or 90 mins and concentrated. The residue was dissolvedin water and slowly acidified to pH 2 with dilute hydrochloric acid togive a yellow precipitate which was filtered and dried to give the titlecompound (237 mg, 67%). δH (DMSO d₆) 9.75 (1H, br m), 8.60 (1H, d, 18.7Hz), 8.43 (2H, m), 7.92 (4H, m), 7.62 (1H, m), 7.52 (3H, m), 7.38 (2H,d, J 8.6 Hz), 5.21 (1H, m), 3.57 (2H, q, J 7.1 Hz), 3.55 (2H, q, J 7.1Hz), 3.3 (1H, dd, J 14.1, 4.6 Hz), 3.15 (1H, dd, J 14.1, 10.1 Hz), 1.14(3H, t, J 7.1 Hz); M/Z (ES+, 70V) 536 (MH⁺).

[0428] The compounds of Examples 59 to 64 were prepared frommethyl-(S)-3-(4-aminophenyl)-24N-t-butoxycarbonyl)aminopropanoate andthe appropriate quinazoline in a manner similar to that described forIntermediate 18 and then derivatised in a manner similar to thatdescribed for Examples 56, 57 and 58.

EXAMPLE 59

[0429]Methyl-(S)-3-[4(Quinazolin-4-ylamino)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0430] δH (CDCl₃) 8.73 (1H, s), 8.0 (1H, d, J 8.5 Hz), 7.91 (1H, d, J8.3 Hz), 7.83-7.54 (6H, m), 7.15 (2H, d, J 8.5 Hz), 5.41 (1H, br s), 3.8(3H, s), 3.70-3.35 (4H, br m), 3.35-3.15 (2H, m), 1.23 (6H, t, J 7.2Hz); m/z (ES⁺, 70V) 474 (MH⁺).

EXAMPLE 60

[0431](S)-3-[4-(Quinazolin-4-ylamino)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0432] δH (d₆ DMSO, 390K) 8.62 (1H, s), 8.55 (1H, d, 18.8 Hz), 7.90-7.82(5H, m), 7.66-7.62 (1H, nr m), 7.34 (2H, d, J 8.5 Hz), 7.09 (1H, br s),5.25 (1H, brs), 3.64-3.56 (4H, m), 3.35 (1H, dd, J 14.2, 5.1 Hz), 3.20(1H, dd, J 14.2, 9.1 Hz), 1.23 (6H, t, J 17.15 Hz); m/z (ES⁺, 70V) 460(MH⁺).

EXAMPLE 61

[0433](S)-3-{4-[(6,7-Dimethoxyquinazolin-4-yl)amino]phenyl}-2-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0434] δH (CDCl₃) 9.39 (1H, s), 8.41 (1H, s), 7.81 (1H, s), 7.67 (3H,dd, J 8.5, 3.8 Hz), 7.25 (2H, d, J 8.4 Hz), 7.16 (1H, s), 5.12 (1H, brs), 3.93 (3H, s), 3.91 (3H, s), 3.60-3.40 (4H, m), 3.20-2.90 (2H, m),1.09 (6H, t, J 7.0 Hz); m/z (ES⁺, 70V) 520 (MH⁺).

EXAMPLE 62

[0435](S)-3-{4-{(6,7-Dimethoxyquinazolin-4-yl)amino]phenyl}2-{[2-n-propylamino)-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0436] δH (DMSO) 9.40 (1H, s), 8.42 (1H, s), 7.81 (1H, s), 7.70 (1H, s),7.66 (2H, d, 18.3 Hz), 7.16 (2H, d, J 7.9 Hz), 7.15 (1H, s), 4.82 (1H,br s), 3.93 (3H, s), 3.91 (3H, s), 3.6-2.9 (4H, m), 1.49 (2H, dd, J14.1, 7.0 Hz), 0.86 (3H, t, J 7.3 Hz); m/z (ES⁺, 70V) 506 (MH⁺).

EXAMPLE 63

[0437] Methyl(S)-3-[4-(6-methoxyquinazolin-4-ylamino)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0438] δH (CDCl₃) 8.65 (1H, s), 7.83 (1H, d, J 9.1 Hz), 7.69 (3H, s, d,J 8.0 Hz), 7.45 (1H, dd, 19.2, 2.6 Hz), 7.13 (2H, d, J 8.5 Hz), 5.40(1H, br s), 3.95 (3H, s), 3.79 (3H, s), 3.6-3.41 (4H, br m), 3.48 (1H,dd, J 14.1, 5.5 Hz), 3.22 (1H, dd, J 14.1, 7.0 Hz), 1.29 (6H, t, 17.2Hz); m/z (ES⁺, 70V) 504 (MH⁺).

EXAMPLE 64

[0439](S)-3-[4-(6-Methoxyquinazolin-4-ylamino)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0440] δ_(H) (d₆ DMSO, 370K); 9.35 (1H, br s), 8.40 (1H, s), 7.89 (1H,d, J 2.7 Hz, 7.54 (2H, d, J 8.6 Hz), 7.71 (1H, s), 7.49 (1H, d, J 2.7Hz), 7.28 (2H, d, J 8.5 Hz), 7.15 (1H, br s), 5.14 (1H, br s), 3.97 (3H,s), 3.42-3.6 (4H, m), 3.28 (1H, dd, J 14.1, 4.9 Hz), 3.60-3.42 (4H, m),3.28 (1H, dd, J 14.1, 4.9 Hz), 3.14 (1H, dd, J 14.1, 9.2 Hz), 1.16 (6H,t, 17.1 Hz); m/z (ES⁺, 70V) 490 (MH⁺)

[0441] The compounds of Examples 65 to 68 were prepared fromIntermediate 32 in a manner similar to that described for Examples 56 to58.

EXAMPLE 65

[0442] Methyl3-{4-[(6,7-dimethyoxy-4-quinazolinyl)amino]-3-methoxyphenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0443] δH (DMSO d₆, 390K) 8.54 (1H, br s), 8.38 (1H, s,), 7.75 (1H, d, J7.8 Hz), 7.66 (1H, s), 7.34 (1H, br d J 8.5 Hz), 7.25 (1H, br s), 7.20(1H, s), 6.97 (1H, d, J 1.9 Hz), 6.85 (1H, br s), 5.15-5.09 (1H, m),3.97 (3H, s), 3.96 (3H, s), 3.78 (3H, s), 3.76 (3H, s), 3.52-3.47 (2H,m), 3.26 (1H, dd, J 14.1, 5.6 Hz), 3.12 (1H, dd, J 14.1, 8.0 Hz), 1.59(2H, sext, J 7.2 Hz), 0.93 (3H, t, J 7.4 Hz); m/z (ES⁺ 70V) 550 (MH⁺).

EXAMPLE 66

[0444] 3-{4-[(6,7-Dimethoxy-4-quinazolinyl)amino]-3-methoxyphenyl}2-(2propylamino-3,4-dioxocyclobut-1-enylamino)propanoic acid

[0445] δH (DMSO d₆, 390K) 8.38 (1H, s), 7.73 (1H, d, J 8.0 Hz), 7.66(1H, s), 7.27 (1H, br s), 7.20 (1H, s), 6.99 (1H, d, J 1.18 Hz), 6.87(1H, dd, J 8.0, 1.9 Hz), 5.02 (1H, m), 3.97 (3H, s), 3.96 (3H, s), 3.83(3H, s), 3.49 (2H, q, J 6.3 Hz), 3.27 (1H, dd, J 14.1, 5.4 Hz), 3.11(1H, dd, J 14.1, 7.8 Hz), 1.59 (2H, sext. J 76.2 Hz), 0.93 (3H, t, 17.4Hz); m/z (ES⁺, 70V) 536 (MH⁺.

EXAMPLE 67

[0446] Methyl3-{4-[(6,7-dimethoxy-4-quinazolinyl)amino]-3-methoxyohenyl}-2(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoateδH (DMSO d₆, 390K) 8.33 (1H, br s), 8.15 (1H, s), 7.51 (1H, d, 18.1 Hz),7.44 (1H, s), 6.98 (1H, s), 6.92 (1H, d, 19.0 Hz), 6.82 (1H, d, J 1.Hz), 6.69 (1H, dd, J 8.0, 1.9 Hz), 5.15-5.09 (1H, m), 3.76 (3H, s), 3.75(3H, s), 3.61 (3H, s), 3.55 (3H, s), 3.35 (4H, q, J 71 Hz), 3.11 (1H,dd, J 14.2, 51 Hz), 2.94 (1H, dd, J 14.2, 9.5 Hz), 0.96 (6H, t, J 7.1Hz); m/z (ES⁺, 70V) 564 (MH⁺).

EXAMPLE 68

[0447]3-{4[(6,7-Dimethoxy-4-quinazolinyl)amino]-3-methoxyphenyl}-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0448] δH (DMSO d₆, 390K), 8.49 (1H, s,), 7.97 (1H, br s), 7.51-7.49(1H, m), 7.38 (1H, s), 7.21 (1H, br d), 7.12 (1H, s), 6.96 (1H, dd, J7.9, 1.3 Hz), 5.28-5.25 (1H, m), 4.00 (6H, s), 3.81 (3H, s), 3.58 (1H,q, J 7.1 Hz), 3.35 (1H, dd, J 14.2, 4.8 Hz), 3.20 (1H, dd, J 14.2, 9.7Hz), 1.18 (6H, t, J 7.1 Hz); m/z (ES⁺, 70V) 550 (MH⁺).

EXAMPLE 69

[0449] Methyl(S)-3-{4-[(6,7-dimethoxy4-quinazolinyl)amino]phenyl}-2-(2-tertbutyl-3,4-dioxocyclobut-1-enylamino)propanoate

[0450] A mixture ofmethyl-(S)₄-[(6,7-dimethoxy-4-quinazolinyl)amino]phenyl}-2-aminopropanoate (332 mg, 0.869 mmol) and Intermediate 4 (171 mg, 0.87 mmol)in MeOH (10 ml) was heated at reflux for 5 days. The solvent was removedin vacuo. The residue was dissolved in DCM, washed with dil. HCl, dried(Na₂SO₄) and concentrated in vacuo. Column chromatography (S)O₂;MeOH/DCM, 7:93) gave the title compound as a brown glass (275 mg). δH(DMSO d₆) 9.39 (1H, s), 8.62 (1H, br d), 8.40 (1H, d, J 1.2 Hz), 7.81 (1h, s), 7.69-7.65 (2H, m), 7.22 (2H, d, 8.5 Hz), 5.08 (1H, m), 3.94 (3H,s), 3.91 (3H, s), 3.74 (3H, s), 3.30 (1H, m), 3.02 (1H, dd, J 13.5, 11.2Hz), 1.22 (9H, s); m/z (ES⁺, 70V) 519 (MH⁺).

EXAMPLE 70

[0451]Ethyl-(S)-3-{4-[(3-chloro-6,7-dimethoxy4-quinazolinyl)amino]phenyl}-2-[(2-isopropoxy-3,4-dioxocyclobut-1-enylamino)propanoate

[0452] Prepared in a similar manner to the compound of Example 56 fromthe Intermediate 45. δH (CD₃OD) 7.73 (2H, d, J 8.6 Hz), 7.73 (1H, s),7.27 (2H, d, J 8.6 Hz), 7.06 (1H, s), 5.28 (1H, m), 5.07 and 4.62 (1H,br), 4.23 (2H, q), 4.00 (3H, s), 3.97 (3H, s), 3.35 (1H, m) 3.05 (1H,m), 1.40 (6H, d, J 6.2 Hz), 1.30 (3H, t, J 7.3 Hz).

EXAMPLE 71

[0453]Ethyl-(S)-3-{4-{(3-chloro-6,7-dimethoxy4-quinazolinyl)amino[phenyl)-2-E(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino]propanoate

[0454] Prepared in a similar manner to the compound of the Example 57from the compound of Example 70. δ_(H) (CD₃OD) 7.72 (1H, s), 7.70 (2H,d, J 8.6 Hz), 7.29 (2H, d, 18.6 Hz), 7.04 (1H, s), δ 33 (1H, dd), 4.25(2H, q, J 7.1 Hz), 3.99 (3H, s), 3.96 (3H, s), 3.58 (4H, br), 3.44 (1H,dd), 3.10 (1H, dd), 1.30 (3H, t, 17.1 Hz), 1.20 (6H, t, J 7.2 Hz).

EXAMPLE 72

[0455](S)-3-{4-[(3-Chloro-6,7-dimethoxy-4-quinazolinyl)amino]phenyl}-2-[(2-N,N-diethylamino-cyclobut-1-enyl)amino]propanoicacid

[0456] Prepared in a similar manner to the compound of Example 58 fromthe compound of Example 71. δH (d₆ DMSO) 7.86 (1H, s), 7.65 (2H, d, J8.6 Hz), 7.31 (2H, d, J 8.6 Hz), 7.16 (1H, s), 5.15 (1H, m), 3.97 (3H,s), 3.95 (3H, s), 3.53 (4H, m), 3.20 (1H, m), 3.13 (1H, m), 1.50 (6H, t,J 7.1 Hz). m/z (ES⁺) 554 (MH⁺).

EXAMPLE 73

[0457](S)-3-{4-[(6,7-Dimethoxy-4-quinazolinyl)amino]phenyl}-2-(2-t-butyl-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0458] Prepared in a similar manner to the compound of Example 58 fromthe compound of Example 69. δ_(H) (DMSO d₆, 370K) 8.40 (1H, s), 7.94(1H, d, 19.2 Hz), 7.83 (1H, s), 7.58 (2H, d, 18.5 Hz), 7.17 (2H, d, 18.6Hz), 7.14 (1H, s), 4.96-4.90 (1H, m), 3.89 (3H, s), 3.87 (3H, s), 3.22(1H, dd, J 14.1, 4.5 Hz),3.04 (1H, dd, J 14.0, 10.2 Hz), 1.17 (9H, s).m/z (ES⁺, 70V) 505 (MH⁺).

EXAMPLE 74

[0459]Ethyl-(S)-3-(4-[5-methyl-4-quinazolinyl)amino]phenyl)-2-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0460] Intermediate 38 (800 mg, 2.3 mmol) and3,4-diisopropoxy-3-cyclobuten 1,2-dione (453 mg, 1 equiv) were stirredat RT in anhydrous MeOH (5 m) for 17 h. The solvent was removed in vacuoand the residue purified by column chromatography (Silica, 75:25EtOAc-isohexane) to give the title compound. δH (DMSO d6; 350K), 8.70(broad signal), 8.50 (1H, s), 7.60 (4H, m), 7.30 (1H, d, J 7.0 Hz), 7.20(2H, d, 8.4 Hz), 5.20 (1H, m). 5.60 (1H, broad s), 4.20 (2H, m), 3.20(1H, m), 3.10 (1H, m), 3.00 (3H, s), 1.30 (6H, d, J 6.2 Hz), 1.20 (3H,m); m/z (ES⁺, 70V) 489 (MH⁺).

EXAMPLE 75

[0461] Ethyl(S)-3(4-[(5-methyl4:quinazolinyl)amino]phenyl)-2-[2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0462] The compound of Example 74 (250 mg, 0.5 mmol) andN,N-diethylamine were stirred at RT in anhydrous (5 ml) MeOH for 17 h.The solvent was removed in vacuo and the residue purified by columnchromatography (Silica; EtOAc to 95% EtOAc: 5% MeOH) to isolate thetitle compound (200 mg) as an off-white solid. δH (DMSO d₆), 8.60 (1H,s), 8.50 (1H, s), 7.80 (1H, d, J 9.1 Hz), 7.60 (4H, m), 7.40 (1H, d, J7,0 Hz), 7.20 (2H, d, 6.8 Hz), 5.20 (1H, m), 4.00 (1H, m), 3.70 (3H, s),3.50 (4H, broad signal), 3.20 (1H, m), 3.00 (1H, m), 2.90 (3H, s), 1.20(6H, m). m/z (ES⁺, 70V) 488 (MH⁺).

EXAMPLE 76

[0463](S)-3-(4-[(5-Methyl4-quinazolinyl)amino]phenyl)-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid

[0464] The compound of Example 75 (190 mg, 0.38 mmol) and lithiumhydroxide monohydrate (19 mg) were stirred in a solvent mixture of MeOH(3 ml), THF (1.5 ml), and water (1.5 ml) for 17 h. The solvent wasremoved in vacuo and the residue dissolved in water, the solutionneutralised with HCl, concentrated in vacuo and the residue purified bycolumn chromatography (Silica; 200:20:3:2 DCM:MeOH:AcOH:H₂O) to isolatethe title compound as yellow solid. δH (DMSO d₆, 350K), 8.50 (1H, broadsignal), 7.70 (3H, broad signal), 7.40-7.20 (3H, m), 5.10 (1H, m), 3.6(4H, m), 3.30 (1H, m), 3.10 (1H, m), 1.10 (6H, t, J 7.2 Hz), m/z (ES⁺,70V) 474 (MH⁺).

[0465] Also prepared in a similar manner to that described for Examples75 and 76 from the compound of Example 74 were the compounds of Examples77 and 78:

EXAMPLE 77

[0466] Methyl(S)-3-(4-[(5-methyl-4-quinazolinyl)amino]phenyl)-2-[(2-N,N-di-n-propylamine-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0467] δH (DMSO d₆, 350K), 8.50 (1H, broad s), 7.80-7.60 (4H, m), 7.40(1H, m), 7.30 (2H, m), 5.30 (1H, m), 3.70 (3H, s), 3.50 (4H, m), 3.40(1H, m), 3.20 (1H, m), 3.10 (3H, s), 1.60 (4H, m), 0.90 (6H, t, J 7.4Hz), m/z (ES⁺, 70V) 516 (MH⁺)

EXAMPLE 78

[0468](S(-3-(4-[(5-Methyl-4-quinazolinyl)amino]phenyl)-2-[(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid δH (DMSO d₆, 350K), 8.70 (1H, s), 7.80 (2H, m), 7.70-7.40 (3H, m),7.40 (2H, d, J 8.5 Hz), 5.20 (1H, m), 3.60 (4H, m), 3.40 (1H, m), 3.20(1H, m), 3.00 (3H, s), 1.50 (4H, m), 0.90 (6H, t, J 7.3 Hz), ml/z (ES⁺,70V) 502 (MH⁺).

EXAMPLE 79

[0469]Ethyl-(S)-3-(4-([6-(trifluoromethoxy)-4-quinazolinyl]amino)phenyl]-2-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0470] Prepared in a similar manner to Example 74 from the compound ofIntermediate 43. δ_(H) (DMSO d₆, 300K), 9.20-9.00 (1H, m), 8.70 (2H, m),8.00 (2H, m), 7.80 (2H, m), 7.30 (2H, m), 5.20 (1H, m), 5.00 (1H, m),4.50 and 4.20 (1H, 2, sets m), 3.80 (3H, m), 3.30 (1H, m), 3.00 (1H, m),1.30 (6H, m), m/z (ES⁺, 70V) 545 (MH⁺).

EXAMPLE 80

[0471] Methyl(S)-3-(4([6-(trifluoromethoxy)-4-quinazolinyl]amino)phenyl)-2-[(2-N,N-diethylamino-3,4-dioxocyclbut-1-enyl)amino]propanoate

[0472] Prepared from the compound of Example 79 in a similar manner tothat described for Example 75. δ_(H) (CD₃OD), 8.50 (1H, s), 8.40 (1H,s), 7.90 (1H, d, 19.1 Hz), 7.80 (1H, m), 7.70 (2H, d, J 8.6 Hz), 7.30(2H, d, J 8.6 Hz), 5.50 (1H, m), 3.80 (3H, s), 3.60 (4H, broad signal),3.50 (1H, m), 3.10 (1H, m), 1.20 (6H, t, J 7.2 Hz), m/z (ES⁺, 70V) 544(MH⁺).

EXAMPLE 81

[0473](S)-3-(4-([6-(Trifluoromethoxy)-4-quinazolinyl]amino)phenyl)-2-[(2-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid

[0474] Prepared from Example 80 in a similar manner to that describedfor Example 76. δH (DMSO, 340K), 9.80 (bd s), 8.60 (2H, m), 8.00-7.70(4H, m), 7.30 (2H, d), 5.20 (1H, m), 3.50 (4H, m), 3.30 (1H, m), 3.10(1H, m), 1 20 (6H, t, 17.1 Hz) m/z (ES⁺, 70V) 544 (MH⁺).

EXAMPLE 82

[0475]Methyl-(S)-3-(4([6-(trifluoromethoxy)-4-quinazolinyl]amino)phenyl)-2-[(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0476] Prepared from the compound of Example 79 in a similar manner tothat described for Example 75. δH (CD₃OD), 8.50 (1H, s), 8.40 (1H, broadsignal), 7.90 (1H, d, 9.2 Hz), 7.85-7.70 (3H, m), 7.30 (1H, d, 18.5 Hz),5.40 (1H, m), 3.80 (3H, s), 3.60 (5H, broad signal), 3.10 (1H, m), 1.60(4H, m), 0.90 (3H, t, J 7.4 Hz), m/z (ES⁺, 70V) 586 (MH⁺).

EXAMPLE 83

[0477](S)-3-(4-([6-(Trifluoromethoxy)-4-quinazolinyl]amino)phenyl)-2-[(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid

[0478] Prepared from the compound of Example 82 in a similar manner tothat described for Example 76. δH (DMSO d₆, 350K), 9.70 (broad signal),8.60 (2H, m), 7.90 (1H, d, J 9.2 Hz), 7.70 (3H, m), 7.30 (2H, d, J 8.0Hz), 5.20 (1H, m), 3.50 (4H, m), 3.30 (1H, m), 3.200 (1H, m), 1.60 (4H,m), 0.90 (3H, t, J 7.4 Hz), m/z (ES⁺, 70V) 572 (MH⁺).

[0479] The compounds of Examples 84 to 89 were prepared in a similarmanner to that described for the preparation of Intermediate 8 fromN-BOC-L-tyrosine methyl ester and the appropriate quinazoline and thenderivatised in a manner similar to that described for Examples 56 to 58.

EXAMPLE 84

[0480] Methyl(S)-3-[4-(6,7-dimethoxyquinazolin-4-yloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoate

[0481] δ_(H) (CDCl₃) 8.57 (1H, s), 7.54 (1H, s), 7.33 (1H, s), 7.25-7.17(5H, m), 5.55-4.9 (1H, m), 4.07 (6H, s), 3.83 (3H, s), 3.55-3.4 (5H, m),3.31 (1H, dd, J 9.0, 5.5 Hz), 1.25 (6H, t, 17.2 Hz), m/z (ES⁺, 70V) 535(MH⁺).

EXAMPLE 85

[0482](S)-3-{4-[(6,7-Dimethoxyquinazolin-4-yl)oxy]phenyl}-2-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0483] δH (d₆ DMSO, 370K) 8.58 (1H, s), 7.64 (1H, s), 7.44-7.40 (3H, m),7.28 (2H, d, J 8.5 Hz), 5.23 (1H, br s), 4.06 (3H, s), 4.04 (3H, s),3.66-3.56 (4H, m), 3.39 (1H, dd, J 14.1, 4.6 Hz), 3.21 (1H, dd, J 14.1,9.6 Hz). 1.22 (6H, t, J 7.1 Hz), m/z (ES⁺, 70V) 521 (MH⁺),

EXAMPLE 86

[0484] Methyl(S)-3-[4-(6-methoxyquinazolin-4-yloxy)phenyl]-2-(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoate.

[0485] δ_(H) (CDCl₃) 8.62 (1H, s), 7.95 (2H, d, 9.0 Hz), 7.59-7.54 (2H,m), 7.26-7.18 (3H, m), 5.47-5.42 (1H, m), 5.30 (1H, d, J 8.4 Hz), 3.99(3H, s), 3.83 (3H, s), 3.60-3.10 (6H, m), 1.67-1.60 (4H, m), 0.92 (6H,t, J 7.4 Hz); m/z (ES⁺, 70V) 533 (MH⁺).

EXAMPLE 87

[0486](S)-3-[4-(6-Methoxyquinazolin-4-yloxy)phenyl]-2-(2-N,N-di-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0487] (d₆ DMSO) 8.57 (1H, s), 7.92 (1H, d, J 10.0 Hz), 7.65 (2H, d, J7.3 Hz), 7.37 (2H, d, J 8.7 Hz), 7.24 (2H, d, J 8.6 Hz), 7.09 (1H, brs), 5.13 (1H, br s), 3.98 (3H, s), 3.59-3.39 (4H, m), 3.35 (1H, dd, J14.5, 5.0 Hz), 3.17 (1H, dd, J 14.1, 9.4 Hz), 1.67-1.50 (4H, m), 0.87(6H, t, J 7.3 Hz); m/z (ES⁺, 70V) 519 (MH⁺)

EXAMPLE 88

[0488](S)-3-[4-(6-Methoxyquinazolin-4-yloxy)phenyl]-2-(2-n-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid δ_(H) (de DMSO, 370K) 8.59 (1H, s), 7.92 (1H, d, J 9.8Jz), 7.65(2H, d, J 7.3 Hz), 7.35-7.24 (5H, m), 4.97 (1H, br s), 3.99 (3H, s),3.50 (2H, t, J 6.3 Hz), 3.29 (1H, dd, J 14.0, 5.4 Hz), 3.13 (1H, dd, J14.1, 7.4 Hz), 1.58 (2H, dd, J 14.2, 7.1 Hz), 0.92 (3H, t, J 7.4 Hz);m/z (ES⁺, 70V) 477 (MH⁺).

EXAMPLE 89

[0489](S)-3-[4-(6-Methoxyquinazolin-4-yloxy)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid δ_(H) (DMSO, 370K) 8.58 (1H, s), 7.92 (1H, d, J 9.9 Hz), 7.65 (2H,d, J 4.7 Hz), 7.39 (2H, d, 18.5 Hz), 7.25 (2H, d, 18.8 Hz), 5.21 (1H, brs), 3.98 (3H, s), 3.6-3.5 (4H, m), 3.34 (1H, dd, J 14.2, 5.1 Hz), 3.17(1H, dd, J 14.1, 9.9 Hz), 1.17 (6H, t, 17.1 Hz); m/z (ES⁺, 70V) 461(MH⁺).

EXAMPLE 90

[0490](S)-Ethyl-3-[4-(isoquinolin-1-ylamino)phenyl]-2-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0491] A solution of Intermediate 10 (426 mg, 1.27 mmol) and3,4-diisopropoxy-3-cyclobutene-1,2-dione (301 mg, 1.52 mmol) in absoluteethanol (5.0 ml) was stirred at 400 under N₂ for 18 h. The volatileswere removed in vacuo and the residue chromatographed (S)O₂; 25-50%EtOAc/hexane) to afford the title compound as a pale orange foam (585mg, 97%). δH (CDCl₃) 8.04 (1H, d, J 5.8 Hz), 7.98 (1H, d, J 8.4 Hz),7.72 (1H, d, J 7.8 Hz), 7.62 (1H, obscured m), 7.61 (2H, d, J 8.3 Hz),7.52 (1H, app.t, J 7.0 Hz), 7.35 (1H, br s), 7.12-7.08 (3H, m), 6.60,6.03, 5.13 and 4.59 (together 1H, m), 5.32 (1H, m), 4.24 (2H, q, J 7.1Hz), 3.25-3.01 (2H, br m), 1.39 (6H, d, J 6.1 Hz), 1.30 (3H, t, J 7.1Hz); m/z (ES⁺, 70V) 474 (MH⁺).

EXAMPLE 91

[0492] Ethyl (S)-3-[4isoquinolin-1-ylamino)phenyl]2-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoateA solution of the compound of Example 90 (585 mg, 1.24 mmol) anddiethylamine (181 mg, 225 μl, 2.48 mmol) in absolute ethanol (2 ml) washeated at 500 under N₂ for 18 h. The volatiles were removed in vacuoaffording the title compound as a dull orange foam (520 mg). δH (CDCl₃)8.05 (1H, d, 15.8 Hz), 7.96 (1H, d, J 8.3 Hz), 7.75 (1H, d, J 7.6 Hz),7.65 (1H, m), 7.63 (2H, d, J 8.5 Hz), 7.55 (1H, app.t, J 7.0 Hz), 7.23(1H, br s), 7.11 (1H, m), 7.10 (2H, d, J 8.5 Hz), 5.39 (1H, narrow m),4.25 (2H, q, J 7.1 Hz), 3.65-3.35 (4H, br m), 1.32 (3H, t, J 7.1 Hz),1.22 (6H, t, J 7.2 Hz); m/z (ES⁺, 70V) 487 (MH⁺).

EXAMPLE 92

[0493](S)-3-[4-(Isoquinolin-1-ylamino)phenyl]-2-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0494] A solution of Example 91 (510 mg, 1.05 mmol) and LiOH.H₂O (53 mg,1.26 mmol) in water (8 ml) and dioxan (8 ml) was stirred at roomtemperatue for 1.5 h. Several drops of AcOH were added and the volatileswere removed in vacuo. The residue was chromatographed [silica, DCM(200-120), MeOH (20), AcOH (3), H₂O(2)]. Freeze-drying from aqueous MeOHafforded the title compound as a pale yellow amorphous solid (230 mg,48%). δH (d⁶ DMSO) 9.07 (1H, br s), 8.49 (1H, d, 18.3 Hz), 7.95 (1H, d,J 5.7H), 7.79-7.75 (3H, m's), 7.70-7.64 (2H, m's), 7.58 (1H, td, J 8.3,1.3 Hz), 7.20 (2H, d, 18.4 Hz), 7.13 (1H, d, 15.6 Hz), 5.11 (1H, m),3.65-3.38 (4H, br m), 3.22 (1H, dd, J 13.9, 4.0 Hz), 2.99 (1H, dd, J13.9, 11.0 Hz) and 1.09 (6H, t, J 7.0 Hz); m/z (ES⁺, 70V) 459 (MH⁺).

EXAMPLE 93

[0495] Ethyl(3S)-3-{4-[(tert-Butoxycarbonyl)amino]phenyl}-3-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0496] Intermediate 12 (190 mg, 0.62 mmol) in MeOH was treated with3,4-diisopropoxy-3-cyclobutene-1,2-dione (122 mg) andN-methyl-morpholine (0.1 ml) and stirred at RT for 16 h. The solvent wasremoved and the product purified by column chromatography(S)O₂;CH₂Cl₂/MeOH 20:1) to give the title compound (176 mg, 64%) as awhite foamy solid. δH (DMSO) 7.41 (2H, d, J 8.6 Hz), 7.24 (2H, d, J 8.6Hz, 5.29 (1H, m), 5.25 (1H, septet, J 6.2 Hz), 4.06 (2H, q, J 7.1 Hz),2.99 (1H, dd, J 15.8, 8.8 Hz), 2.86 (11H, dd, J 15.8, 6.0 Hz), 1.40 (3H,d, J 6.2 Hz), 1.36 (3H, d, 16.2 Hz), 1.16 (3H, t, J 7.1 Hz); m/z (ESI,70V) 469 (MNa⁺).

EXAMPLE 94

[0497] Ethyl(3S)-3-(4-aminophenyl)-3-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0498] The compound of Example 93 (176 mg, 0.39 mmol) was dissolved inEtOAc (10 ml) and HCl gas was bubbled through. The reaction mixture wasstirred for 2 h and the solvent removed to give the title compound (130mg, 0.34 mmol, 87%) as an oil. δH (DMSO 360K) 7.38 (2H, d, J 8.5 Hz),7.21 (2H, d, J 8.5 Hz), 5.30 (1H, br m), 5.25 (1H, septet, J 6.2 Hz),4.08 (2H, q, J 7.1 Hz), 2.99 (11H, dd, J 15.8, 8.8 Hz), 2.85 (1H, dd, J15.8, 6.0 Hz), 1.40 (3H, d, J 6.2 Hz), 1.36 (3H, d, J 6.2 Hz), 1.15 (3H,t, J 7.1 Hz).

EXAMPLE 95

[0499] Ethyl(3S)-3-{4-(3,5-dichloro4-pyridylcarboxamido)phenyl}-3-[(2-isopropoxy-3,4-dioxocyclobut-1-enyl)amino]propanoate

[0500] The compound of Example 94 (max 2 mmol) was dissolved in DCM (5ml) and N-methylmorpholine (1 equiv) and cooled to 0°.3,5-dichloroisonicotinoyl chloride (463 mg) was added and the reactionmixture stirred at RT for 16 h then quenched with sodium bicarbonatesolution. The organic layer was washed with dilute hydrochloric acid,water, dried (Na₂SO₄), filtered and the solvent removed. The product waspurified by column chromatography (S)O₂; CH₂Cl₂/MeOH 20:1) to give thetitle compound (636 mg, 61%) as an oil. δH (DMSO, 390K) 10.47 (1H, brs), 8.69 (2H, s), 7.62 (2H, d, 18.4 Hz), 7.39 (2H, d, J 8.4 Hz), 5.38(1H, m), 5.25 (1H, septet, J 6.1 Hz), 4.10 (2H, q, 17.1 Hz), 3.05 (1H,dd, J 15.8, 8.6 Hz), 1.42 (3H, d, J 6.1 Hz), 1.38 (3H, d, J 6.1 Hz),1.18 (3H, t, J 7.1 Hz);; m/z (ES⁺, 70V) 522 (MH⁺)

EXAMPLE 96

[0501] Ethyl(3S)-3d4-(3,5-dichloropyrid-4-ylcarboxamido)phenyl}-3-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoate

[0502] The compound of Example 95 (318 mg, 0.61 mmol) was dissolved inMeOH (5 ml) and diethylamine (0.13 ml). The solution was stirred for 16h to give a white precipitate which was isolated by filtration and driedto give the title compound (247 mg, 78%) as a white solid. δH (DMSO,370K) 10.93 (1H, br s), 8.78 (2H, s), 7.61 (2H, d, J 9.0 Hz), 7.41 (2H,d, J 9.0 Hz), 5.83 (1H, m), 3.59 (3H, s), 3.53 (4H, br m), 3.08 (1H, dd,J 16.0, 9.0 Hz), 2.95 (1H, dd, J 16.0, 6.0 Hz), 1.10 (6H, t, 16.0 Hz);m/z (ES⁺, 70V) 521 (MH⁺).

EXAMPLE 97

[0503](3S)-3-{4-(3,5-Dichloropyrid-4-yl-carboxamido)phenyl}-3-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0504] The compound of Example 96 (235 mg, 0.45 mmol) was dissolved inTHF (5 ml) and water (5 ml) and lithium hydroxide (21 mg) added. Thesolution was stirred at RT for 3 h and the solvent removed in vacuo. Theresidue was dissolved in water (10 ml) and acidified to pH 2 with dil.HCl to give a white precipitate (198 mg, 0.39 mmol, 87%) which wasfiltered and dried to afford the title compound. δH (DMSO, 390K) 10.43(1H, br s), 8.69 (2H, s), 7.60 (2H, d, J 8.5 Hz), 7.45 (2H, d, J 8.5Hz), 7.29 (1H, br s), 5.82 (1H, m), 3.60 (2H, q, J 7.0 Hz), 3.58 (2H, q,J 7.0 Hz), 3.02 (1H, dd, J 15.8, 8.2 Hz,), 2.90 (1H, dd, J 15.8, 6.1Hz), 1.20 (6H, t, J 7.0 Hz); m/z (ES⁺, 70V) 507 (MH⁺). Analysis bychiral HPLC on Chirobiotic T column eluting with MeOH/0.6%HOAc gavesingle peak eluting at 5.58 minutes.

EXAMPLE 98

[0505](3R)-3-{4-(3,5-Dichloropyrid4-ylcarboxamido)phenyl-3-{[2-N,N-(diethylamino)-3,4-dioxo-1-cyclobutenyl]amino}propanoicacid

[0506] This was prepared by the same route as the (S)-enantiomer Example97 using the appropriate chiral amine. Analysis by chiral HPLC onChirobiotic T column eluting with MeOH/0.6%HOAc gave single peak elutingat 6.54 minutes.

EXAMPLE 99

[0507] Methyl(3R)-3-[(2-Isopropoxy-3,4-dioxocyclobut-1-enyl)amino]-3-{4-[(6,7-dimethoxy4-quinazolinyl)oxy]phenyl}propanoate

[0508] Intermediate 16 (580 mg, 1.38 mmol) was dissolved in MeOH (6 ml)and DIPEA (0.53 ml) and 3,4-diisopropoxy-3-cyclobuten-1,2-dione (300 mg)added. The solution was stirred for 16 h and the solvent removed. Theresidue was purified by column chromatography (S)O₂; CH₂Cl₂/MeOH 50:1)to give the title compound (539 mg, 75%) as a yellow oil. δH (DMSO,350K) 8.99 (1H, br m), 8.54 (1H, s), 7.57 (1H, s), 7.49 (2H, d, J 8.6Hz), 7.38 (1H, s), 7.32 (2H, d, 8.6 Hz), 5.40 (11H, m), 5.27 (1H,septet, J 6.2 Hz), 4.01 (3H, s), 3.98 (3H, s), 3.64 (3H, s), 3.10 (1H,dd, J 16.1, 5.8 Hz,), 2.97 (1H, dd, J 16.1, 5.8 Hz), 1.42 (3H, d, J 6.2Hz), 1.38 (3H, d, J 6.2 Hz); m/z (ES⁺, 70V) 522 (MH⁺).

EXAMPLE 100

[0509] Methyl(3R)-3-{[2-N,N-diethylamino-3,4-dioxo-1-cyclobutenyl]amino}-3-{4-[(6,7-dimethoxy4-quinazolinyl)oxy]phenyl}propanoate

[0510] The compound of Example 99 (265 mg, 0.51 mmol) was dissolved inMeOH (3 ml) and diethylamine added (0.1 ml). The solution was stirredfor 16 h giving a white precipitate. The precipitate was filtered anddried to give the title compound (177 mg, 65%) as a white solid. δH(DMSO, 370K) 8.55 (1H, s), 7.59 (1H, s), 7.54 (2H, d, J 8.5 Hz), 7.32(1H, s), 7.30 (2H, d, J 8.5 Hz, 5.94 (1H, m), 4.02 (3H, s), 3.99 (3H,s), 3.64 (3H, s), 3.60 (4H, septet, J 7.1 Hz), 3.15 (1H, dd, J 15.7, 8.9Hz), 3.03 (1H, dd, J 15.7, 5.9 Hz), 1.19 (6H, t, J 7.1 Hz),; m/z (ES⁺,70V) 535 (MH⁺).

EXAMPLE 101

[0511] (R)-3-{[2-N,N-7Diethylamino-3,4-dioxo-1 cyclobutenyl]amino}-3-{4[(6,7-dimethoxy-4-quinazolinyl)oxy]phenyl}propanoic acid

[0512] The compound of Example 100 (170 mg, 0.32 mmol) was dissolved inTHF (2 ml) and water (2 ml) and lithium hydroxide (20 mg) was added. Thesolution was stirred at RT for 3 h and the solvent removed. The residuewas dissolved in water (10 ml) and acidified to pH 2 with dil. HCl togive a white precipitate (42 mg, 25%) which was filtered and dried-δH(DMSO, 400K) 8.56 (1H, s), 7.60 (1H, s), 7.54 (2H, d, J 8.6 Hz), 7.39(1H, s), 7.31 (2H, d, J 8.6 Hz), 5.90 (1H, m), 4.03 (3H, s), 3.99 (3H,s), 3.62 (2H, q, J 7.1 Hz), 3.60 (2H, q, J 7.1 Hz), 3.06 (1H, dd, J15.8, 8.2 Hz), 2.95 (1H, dd, J 5.8, 6.1 Hz), 1.21 (3H, t, J 7.1 Hz),;m/z (ES⁺ 70V) 521 (MH⁺).

EXAMPLE 102

[0513] Ethyl(R)-3-[(2-Isopropoxy-3,4-dioxo-1-cyclobutenyl)amino-3-[4-(2,6-naphthyridin-1-ylamino)phenylpropanoate

[0514] Intermediate 23 (178 mg, 0.53 mmol) was dissolved in MeOH (5 ml)and DIPEA (0.2 ml), treated with 3,4-diisopropoxy-3-cyclobuten-1,2-dione(126 mg) and stirred at RT for 16 h. The solution was concentrated,dissolved in DCM (20 ml), washed with water, dried (Na₂SO₄), filteredand concentrated. The crude product was purified by columnchromatography (S)O₂, CH₂Cl₂/MeOH 50:1) to give the title compound (150mg, 60%) as an oil. (H (DMSO, 370K) 9.21 (1H, s), 9.09 (1H, br s), 8.70(1H, br m), 8.65 (1H, d, J 5.9), 8.33 (1H, d, J 5.9 Hz), 8.16 (1H, d, J5.7 Hz), 7.87 (2H, d, J 8.5 Hz), 7.35 (2H, d, J 8.5 Hz), 7.28 (1H, d, J5.7 Hz), 5.37 (1H, m), 5.27 (1H, septet, J 6.2 Hz), 4.10 (2H, qd, J 7.1,0.4 Hz), 3.05 (1H, dd, J 15.8, 8.9Ht), 2.93 (1H, dd, J 15.8, 5.9), 1.43(3H, d, J 6.2 Hz), 1.39 (3H, d, J 6.2H)), 1.18 (3H, t, 17.1 Hz); m/z(ES⁺, 70V) 475 (MH⁺).

EXAMPLE 103

[0515] Methyl(3R)-3-[(2-N,N-Diethylamino-3,4-dioxo-1-cyclobutenyl)amino-3-[4-(2,6-naphthyridin-1-ylamino)phenyl]propanoate

[0516] The compound of Example 102 (145 mg, 0.3 mmol) in MeOH (2 ml) wastreated with diethylamine (0.07 ml) and stirred at RT for 16 h. Thesolvent was removed and the residue was purified by columnchromatography (S)O₂, CH₂Cl₂/MeOH 100:1) to give the title compound (140mg, 98%) as a yellow oil. δH (DMSO, 370K) 9.21 (1H, s), 9.08 (1H, br s),8.65 (1H, d, J 5.9 Hz), 8.33 (1H, d, 15.9 Hz). 8 16 (1H, d, J 5.7 Hz),7.86 (2H, d, J 8.5 Hz), 7.40 (2H, d, J 8.5 Hz), 7.27 (1H, d, J 5.7 Hz),5.87 (1H, m), 3.63 (3H, s), 3.59 (2H, q, J 7.1 Hz), 3.57 (2H, q, J 7.1Hz), 3.12 (1H, dd, J 15.6, 8.8 Hz), 2.99 (1H, dd, J 15.6, 6.0 Hz), 1.21(3H, t, J 7.1 Hz), 1.18 (3H, t, J 7.1 Hz), m/z (ES⁺, 70V) 474 (MH⁺).

EXAMPLE 104

[0517](3R)-3-[(2-N,N-Diethylamino-3,4-dioxo--cyclobutenyl)amino-3-[4-(2,6-naphthyridin-1-ylamino)phenylpropanoicacid

[0518] The compound of Example 103 (140 mg, 0.29 mmol) was dissolved inTHF (1 ml) and water (1 ml) and treated with lithium hydroxide (18 mg).The solution was stirred at RT for 90 mins and concentrated in vacuo.The residue was dissolved in water and slowly acidified to pH 4.5 withdilute HCl acid to give a yellow precipitate which was filtered anddried to give the title compound (60 mg). δH (DMSO, 350K) 9.22 (1H, d, J0.8 Hz), 8.66 (1H, d, J 5.8 Hz), 8.36 (11H, dd, J 5.9, 0.8 Hz), 8.15(1H, d, J 5.8 Hz), 7.84 (2H, d, J 8.5 Hz), 7.40 (2H, d, J 8.5 Hz), 7.29(1H, d, J 5.8 Hz), 5.83 (1H, m), 3.59 (2H, q, J 7.1 Hz), 3.57 (2H, q, J7.1 Hz), 3.02 (1H, dd, J 15.7, 8.8 Hz, 2.90 (1H, dd, J 15.7, 5.9 Hz),1.18 (6H, t, J 7.1 Hz), m/z (ES⁺, 70V) 460 (MH⁺).

[0519] The following derivatised resins were prepared to enable thepreparation of compounds of the invention by solid phase synthesis:

[0520] Resin bound(S)-3-(4-Aminophenyl)-2-(9-fluorenylmethoxy-carbonylamino)propanoic acid(1)

[0521] Paramax Wang resin (Advanced Chemtech, 10 g, 1.0 mmol/g, 10 mmolequivalent) in DMF (150 ml) was treated withN-α-FMOC4-nitro-L-phenylalanine (22 g, 50 mmol), 2,6-dichlorobenzoylchloride (7.0 ml, 50 mmol) and pyridine (4.0 ml, 50 mmol) and themixture agitated under nitrogen at RT for 24 h. The resin was filteredand washed with DMF and DCM then unreacted resin sites were capped with20% acetic anhydride in DMF for 30 mins at RT. The resin was filteredand washed as before then treated with a 1 M solution of stannouschloride dihydrate in DMF (100 ml) at RT for 12 h and washed with DMFand DCM to give the derivatised resin (1).

[0522] Resin bound(S)-3-[4-(3,5-dichloro4-pyridylcarboxamido)phenyl]-2-aminopropanoic acid(2)

[0523] Derivatised resin (1) from the above procedure was swollen in DCM(50 ml) then treated with DIPEA (5.1 ml, 29 mmol) and3,5-dichloropyridine-4-carbonylchloride (6.2 ml, 29 mmol) and agitatedunder nitrogen at RT for 12 h. The resin was washed as before thentreated with a 20% solution of piperidine in DMF (100 ml) for 30 mins atRT followed by thorough washing with DMC and DCM to give the derivatisedresin (2).

[0524] Resin bound(S)-3-[4-(3,5-dichloro-4-pyridylcarboxamido)phenyl]-2-(2-methoxy-3,4-dioxocyclobut-1-enylamino)propanoicacid (3)

[0525] Derivatised resin (2) from the above procedure in DMF (100 ml)was treated with 3,4-dimethoxy-3-cyclobutene-1,2-dione (4.1 g, 29 mmol)for 12 h at 70° then filtered and washed with DMF and DCM to give thederivatised resin (3).

[0526] Resin bound(RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-methoxy-3,4-dioxocyclobut-1-enylamino)propanoicacid (4)

[0527] Derivatised resin (4) was prepared in a similar manner toderivatised resin (3) from(RS)-3-(9-fluorenylmethoxycarbonylamino)-3-(4-nitrophenyl) propanoicacid. The latter was prepared as follows: A cold (O) solution of(RS)-3-Amino-3-(4-nitrophenyl)propanoic acid [D. M. Kalvin and R. W.Woodward; J. Org. Chem. (1985) 50, 2259] (3.2 g, 15 mmol) in 10% aqueoussodium carbonate (60 ml) and 1,4-dioxane (30 ml) was treatedportion-wise with 9-fluorenylmethoxycarbonyl-N-hydroxysuccinimide (5.6g, 17 mmol) in 1,4-dioxane (15 ml) and the mixture stirred at RT for 12h. The mixture was poured into water (300 ml) and the aqueous phasewashed 3 times with Et₂O. The aqueous layer was then acidified withsolid citric acid and extracted into Et₂O. The combined organic layerswere dried (MgSO₄) and evaporated to a yellow oil. then triturated fromhexane and EtOAc to afford(RS)-3-(9-fluorenylmethoxy-carbonylamino)-3-(4-nitrophenyl)propanoicacid as a yellow solid (1.8 g); m/z (ES⁺, 70V) 432.

[0528] Resin bound(S)-3d4-Aminophenyl)-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoic acid (5)

[0529] Paramax Wang resin (Advanced Chemtech, 10 g, 1.0 mmol/g, 10 mmolequivalent) in DMF(150 ml) was treated withN-α-FMOC4-nitro-L-phenylalanine (22 g, 50 mmol), 2,6-dichlorobenzoylchloride (7.0 ml, 50 mmol) and pyridine (4.0 ml, 50 mmol) and themixture agitated under nitrogen at RT for 24 h. The resin was filteredand washed with DMF and DCM then unreacted resin sites were capped with20% acetic anhydride in DMF for 30 mins at RT. The resin was filteredand washed as before. A portion (4 g) was treated with a 20% solution ofpiperidine in DMF (100m) for 30 mins at RT then filtered and washed withDMF and DCM. The resin was treated with3,4-dimethoxy-3-cyclobutene-1,2-dione (1.9 g, 13.4 mmol) in DMF (50 ml)for 12 h at 70° C. then filtered and washed with DMF and DCM. The resinwas swollen in DCM (10 ml) and EtOH (40 ml) and treated with propylamine(1.6 ml, 19.2 mmol). The solution was agitated for 12 h at RT thenfiltered and washed thoroughly with DCM. The resin was treated with a 1M solution of stannous chloride dihydrate in DMF (50 ml) at RT for 8 hthen washed with DMF and DCM to give derivatised resin (5).

[0530] Resin bound diethylphosphono-α-diazoacetate (6)

[0531] Wang resin (Advanced Chem tech, 1.0 g, 0.7 mmol/g, 0.7 mmolequivalent) was treated with diethyl-phosphonoacetate (0.68 g, 3.5mmol), N,N′-diisopropylcarbodiimide 0.55 ml, 3.5 mmol) and4-N,N-dimethylamino-pyridine (0.09 g, 0.7 mmol), in DCM (5.0m). Themixture was agitated at ambient temperature for 16 h. The resin wasfiltered and washed with DMF, MeOH and DCM. The resulting resin (1.0 g)was treated with 4-acet-amidobenzenesulfonyl azide (0.43 g, 1.86 mmol)and diazabicyclo-undec-7-ene (0.09 g, 0.62 mmol) in acetonitrile atambient temperature for 16 h. The resin was washed with DMF, MeOH andDCM to give derivatised resin (6) [FTIR (ATR) ν_(max) 2132 cm⁻¹].

[0532] Resin bound(S)-3-[4-(1-isoquinolylamino)phenyl]-2-(9-fluorenylmethoxycarbonylamino)propanoicacid (7)

[0533] Wang resin (Advanced Chemtech, 3.0 g, 0.7 mmol/g, 2.1 mmolequivalent) in DMF (50 ml) was treated with($)-3-[4-(1-isoquinolylamino)phenyl]-2-(9-fluorenylmethoxycarbonylamino)propanoicacid (3.3 g, 6.3 mmol), 2,6-dichlorobenzoyl chloride (1.5 ml, 10.5 mmol)and pyridine (0.8 ml, 10.5 mmol) and the mixture agitated under nitrogenat RT for 24 h. The resin was filtered and washed with DMF and DCM thenunreacted resin sites were capped with 20% acetic anhydride in DMF for30 mins at RT. The resin was filtered and washed as before to givederivatised resin (7).

[0534] Resin bound (S)-3-[4-(1isoquinolylamino)phenyl]-2-(2-methoxy-3,4-dioxocyclobut-1-enylaminopropanoicacid (8)

[0535] Derivatised resin (7) from the above procedure was treated with a20% solution of piperidine in DMF (100 ml) for 30 mins at RT followed bythorough washing with DMF and DCM. The resin was treated with3,4-dimethoxy-3-cyclobutene-1,2-dione (4.7 g, 33 mmol) for 12 h at 70°in DMF (50 ml) then filtered and washed as before to give derivatisedresin (8).

[0536] Resin bound(S)-3-(4-benzoylphenyl)-2-(2-methoxy-3,4-dioxocyclobut-1-enyl)aminopropanoicacid (9b

[0537] N-α-FMOC-L-benzoylphenylalanine Wang resin (Advanced Chemtech,400 mg, 0.5 mmol/g, 0.2 mmol equivalent) was treated with a 20% solutionof piperidine in DMF (5 ml) for 30 mins at RT then filtered and washedthoroughly with DMF and DCM. The resin was treated with3,4-dimethoxy-3-cyclobutene-1,2-dione (200 mg, 1.4 mmol) for 12 h at 700in DMF (5 ml) then filtered and washed as before to give derivatisedresin (9).

[0538] HPLC-MS

[0539] HPLC-M& was performed on a Hewlett Packard 1100/MSD ES SingleQuadropole system with diode array detector using either:

[0540] Conditions A: A Luna C18(2) 50×4.6 mm (3am particle size) column,running a gradient of 95% [20 mM ammonium formate, pH 3.5], 5% [0.1%formic acid in acetonitrile] to 10% [20 mM ammonium formate, pH 3.5],90% (0.1% formic acid in acetonitrile] over 3 min, then maintaining themobile phase at that ratio for a further 2 min. Flow rate 0.8 ml/min.;or

[0541] Conditions B: A Luna C18(2) 50×2.0 mm (3 gm) column, running agradient of 95% [0.1% aqueous formic acid], 5% [0.1% formic acid inacetonitrile] to 10% [0.1% aqueous formic acid], 90% [0.1% formic acidin acetonitrile] over 2 min, then maintaining the mobile phase at thatratio for a further 1 min. Flow rate 0.8 ml/min. MS was acquired by APIelectrospray in positive ion mode, at 70V, scanning from 150 to 750 amu.

EXAMPLE 105

[0542](S)-3-4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl1-212-cyclohexylamino-3,4-dioxocyclobut-1-enylamino)ppropanoicacid

[0543] To the derivatised resin (3), (120 mg) was added DCM (0.2 ml),EtOH (0.8 ml) and a 1M solution of cyclohexylamine in DCM (0.5 ml). Thesolution was agitated for 12 h at RT followed by filtration and multiplewashes with DCM. The resin was treated with 60% trifluoroacetic acid inDCM (1.5 ml) for 3 h with agitation and then filtered. The filtrate wasevaporated in vacuo to give the crude product which was purified bypreparative HPLC to afford the title compound (5 mg).

[0544] HPLC-MS (Conditions A) Retention time 3.5 min MH⁺ 531.

[0545] The following compounds of Examples 106 to 179 and 183 to 195were prepared in a similar manner to the compound of Example 105, eachusing the starting material shown. For examples where the amine wasadded as a salt, 1 mol equivalent of DIPEA was also added.

EXAMPLE 106

[0546](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(1-adamantylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0547] 1-Adamantylamine gave the title compound (4 mg)

[0548] HPLC-MS (Conditions A) Retention time 3.9 min MH⁺ 583

EXAMPLE 107

[0549](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-methoxyethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0550] 2-Methoxyethylamine gave the title compound (10 mg)

[0551] HPLC-MS (Conditions A) Retention time 3.1 min MH⁺ 507

EXAMPLE 108

[0552](S)-3-[4-(3.-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3-methoxypropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid 3-Methoxypropylamine gave the title compound (9 mg)

[0553] HPLC-MS (Conditions A) Retention time 3.2 min MH⁺ 521

EXAMPLE 109

[0554](S)-3-[4-(3,5-Dichloro-4-1pyridylcarboxamido)phenyl]-2-[2-[2-thienylmethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0555] 2-(Aminomethyl)thiophene gave the title compound (4 mg)

[0556] HPLC-MS (Conditions A) Retention time 3.4 min MH⁺ 545

EXAMPLE 110

[0557](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(4-morpholinoethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0558] N-(2-Aminoethyl)morpholine gave the title compound (8 mg)

[0559] HPLC-MS (Conditions A) Retention time 2.9 min MH⁺ 562

EXAMPLE 111

[0560](S)-3-[4-(3,5-Dichloro-4-2pyridylcarboxamido)phenyl]-2-[2-(3,4,5-trimethoxybenzylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0561] 3,4,5-Trimethoxybenzylamine gave the title compound (3 mg)

[0562] HPLC-MS (Conditions A) Retention time 3.4 min MH⁺ 629

EXAMPLE 112

[0563](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(1-piperidinoethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0564] 1-(2-Aminoethyl)piperidine gave the title compound (11 mg)

[0565] HPLC-MS (Conditions A) Retention time 2.9 min MH⁺ 560

EXAMPLE 113

[0566](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-[3-(2-oxopyrrolidin-1-yl)propylamino)-3,4-dioxocyclobut-1-enylaminopropanoic acid

[0567] 1-(3-Aminopropyl)-2-pyrrolidinone gave the title compound (12 mg)

[0568] HPLC-MS (Conditions A) Retention time 3.1 min MH⁺ 574

EXAMPLE 114

[0569](S)-3-4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3phenylpropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0570] 3-Phenylpropylamine gave the title compound (8 mg)

[0571] HPLC-MS (Conditions A) Retention time 3.7 min MH⁺ 567

EXAMPLE 115

[0572](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3-(1-imidazolyl)propylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[0573] N-(3-Aminopropyl)imidazole gave the title compound (9 mg)

[0574] HPLC-MS (Conditions A) Retention time 2.8 min MH⁺ 557

EXAMPLE 116

[0575](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-piperponylamino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0576] Piperonylamine gave the title compound (3 mg)

[0577] HPLC-MS (Conditions A) Retention time 3.5 min MH⁺ 583

EXAMPLE 117

[0578](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(1-benzyl-4-piperidinylamino)-3,4-dioxocyclobut-1-enylamino]ppropanoicacid

[0579] 4-Amino-1-benzylpiperidine gave the title compound (12 mg)

[0580] HPLC-MS (Conditions A) Retention time 3.1 min MH⁺ 622

EXAMPLE 118

[0581](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-pyridylmethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0582] 2-(Aminomethyl)pyridine gave the title compound (14 mg)

[0583] HPLC-MS (Conditions A) Retention time 3.2 min MH⁺ 540

EXAMPLE 119

[0584](S)-3-[4-(3,5-Dichloro-4-pyrdylcarboxamido)phenyl]-2-(2-cyclopentylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0585] Cyclopentylamine gave the title compound (8 mg)

[0586] HPLC-MS (Conditions A) Retention time 3.4 min MH⁺ 517

EXAMPLE 120

[0587](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(4-phenylbutylamino)-3,4-dioxocyclobut-1enylamino]propanoic acid

[0588] 4-Phenylbutylamine gave the title compound (4 mg)

[0589] HPLC-MS (Conditions A) Retention time 3.8 min MH⁺ 581

EXAMPLE 121

[0590](s)-3-[4-(3,5-Dichloro-4-1pyridylcarboxamido)phenyl]-2-[2-(3-pyridylmethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0591] 3-(Aminomethyl)pyridine gave the title compound (7 mg)

[0592] HPLC-MS (Conditions A) Retention time 3.0 min MH⁺ 540

EXAMPLE 122

[0593](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3,3-dimethylbutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0594] 3,3-Dimethylbutylamine gave the title compound (7 mg)

[0595] HPLC-MS (Conditions A) Retention time 3.6 min MH⁺ 533

EXAMPLE 123

[0596](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3,4-dichlorobenzylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0597] 3,4-Dichlorobenzylamine gave the title compound (11 mg)

[0598] HPLC-MS (Conditions A) Retention time 3.8 min MH⁺ 607

EXAMPLE 124

[0599](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-(1-piperazinyl)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[0600] N-(2-aminoethyl)piperazine gave the title compound (5 mg)

[0601] HPLC-MS (Conditions A) Retention time 2.8 min MH⁺ 561

EXAMPLE 125

[0602](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-(1-pyrrolidinyl)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[0603] 1-(2-aminoethyl)pyrrolidine gave the title compound (9 mg)

[0604] HPLC-MS (Conditions A) Retention time 2.9 min MH⁺ 546

EXAMPLE 126

[0605](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(3-hydroxypropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0606] 3-Hydroxypropylamine gave the title compound (4 mg)

[0607] HPLC-MS (Conditions A) Retention time 3.0 min MH⁺ 507

EXAMPLE 127

[0608](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[244cyclohexylanilino)3,4-dioxocyclobut-1-enylamino]propanoicacid

[0609] 4-Cyclohexylaniline gave the title compound (3 mg)

[0610] HPLC-MS (Conditions A) Retention time 4.3 min MH⁺ 607

EXAMPLE 128

[0611](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(4-morpholinoanilino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0612] 4-Morpholinoaniline gave the title compound (5 mg)

[0613] HPLC-MS (Conditions A) Retention time 3.4 min MH⁺ 610

EXAMPLE 129

[0614](S)-3-[4:(3,5-Dichloro4-pyridylcarboxamido)phenyl]-242-isopropylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0615] Isopropylamine gave the title compound (2 mg)

[0616] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 491

EXAMPLE 130

[0617](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-242-tert-butylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0618] Tert-butylamine gave the title compound (1 mg)

[0619] HPLC-MS (Conditions B) Retention time 2.39 min MH⁺ 505

EXAMPLE 131

[0620](S)-3-[4-(3,5-Dichloro-4-pyrodylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0621] Propylamine gave the title compound (5 mg)

[0622] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 491

EXAMPLE 132

[0623](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-benzylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0624] Benzylamine gave the title compound (4 mg)

[0625] HPLC-MS (Conditions B) Retention time 2.43 min MH⁺ 539

EXAMPLE 133

[0626](S)-3-[4-(3,5-Dichloro4-pyrodylcarboxamido)phenyl]-2-[-2-(3-(dimethylamino)propylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0627] 3-(Dimethylamino)propylamine gave the title compound (5 mg)

[0628] HPLC-MS (Conditions B) Retention time 1.92 min MH⁺ 534

EXAMPLE 134

[0629](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(3-isopropoxypropylamino]-3,4-dioxocyclobut-1-enylamino]propanoic

[0630] 3-Isopropoxypropylamine gave the title compound (5 mg)

[0631] HPLC-MS (Conditions B) Retention time 2.37 min MH⁺ 549

EXAMPLE 135

[0632](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(3-ethoxypropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0633] 3-Ethoxypropylamine gave the title compound (5 mg)

[0634] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 535

EXAMPLE 136

[0635](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-(3-indolyl)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0636] 2-(3-indolyl)ethylamine gave the title compound (1 mg)

[0637] HPLC-MS (Conditions B) Retention time 2.15 min MH⁺ 592

EXAMPLE 137

[0638](S)-3-[4(3,5-Dichloro-4-pyridylcarboxamido)phenyl-2-(2-cyclobutylamino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0639] Cyclobutylamine gave the title compound (4 mg)

[0640] HPLC-MS (Conditions B) Retention time 2.35 min MH⁺ 503

EXAMPLE 138

[0641](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-cyclopropylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0642] Cyclopropylamine gave the title compound (5 mg)

[0643] HPLC-MS (Conditions B) Retention time 2.26 min MH⁺ 489

EXAMPLE 139

[0644](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(4-(1,2,3-thiadiazol-4-yl)benzylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0645] 4-(1,2,3-Thiadiazol4-yl)benzylamine gave the title compound (5mg)

[0646] HPLC-MS (Conditions B) Retention time 2.46 MH⁺ 623

EXAMPLE 140

[0647](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2(3-nitrobenzylamino)-3,4dioxocyclobut-1-enylamino]propanoicacid

[0648] 3-Nitrobenzylamine hydrochloride gave the title compound (4 mg)

[0649] HPLC-MS (Conditions B) Retention time 2.46 min MH⁺ 584

EXAMPLE 141

[0650](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(4-(methylsulfonyl)benzylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0651] 4-(Methylsulfonyl)benzylamine hydrochloride gave the titlecompound (1 mg)

[0652] HPLC-MS (Conditions B) Retention time 2.31 min MH⁺ 617

EXAMPLE 142

[0653](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-(benzylthio)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0654] 2-(Benzylthio)ethylamine hydrochloride gave the title compound (7mg)

[0655] HPLC-MS (Conditions B) Retention time 2.56 min MH⁺ 599

EXAMPLE 143

[0656](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(2-(4-nitrophenyl)ethylamino)-3,4dioxocyclobut-1-enylamino]propanoicacid

[0657] 2-(4-Nitrophenyl)ethylamine hydrochloride gave the title compound(1 mg)

[0658] HPLC-MS (Conditions B) Retention time 2.47 min MH⁺ 598

EXAMPLE 144

[0659](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(1-piperidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0660] Piperidine gave the title compound (5 mg).

[0661] HPLC-MS (Conditions B) Retention time 2.36 min MH⁺ 517

EXAMPLE 145

[0662](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-morpholino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0663] Morpholine gave the title compound (7 mg)

[0664] HPLC-MS (Conditions B) Retention time 2.24 min MH⁺ 519

EXAMPLE 146

[0665](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-thiomorpholino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0666] Thiomorpholine gave the title compound (1 mg)

[0667] HPLC-MS (Conditions B) Retention time 2.36 min MH⁺ 535

EXAMPLE 147

[0668](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0669] Diethylamine gave the title compound (4 mg)

[0670] HPLC-MS (Conditions B) Retention time 2.34 min MH⁺ 505

EXAMPLE 148

[0671](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0672] Pyrrolidine gave the title compound (2 mg)

[0673] HPLC-MS (Conditions B) Retention time 2.29 min MH⁺ 503

EXAMPLE 149

[0674](S)-3-14-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(4-ethyl-1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0675] 1-Ethylpiperazine gave the title compound (6 mg)

[0676] HPLC-MS (Conditions B) Retention time 1.96 min MH⁺ 546

EXAMPLE 150

[0677](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(4-(hydroxypropyl)-1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0678] 1-Piperazinepropanol gave the title compound (6 mg)

[0679] HPLC-MS (Conditions B) Retention time 1.94 min MH⁺ 576

EXAMPLE 151

[0680](S)-3-[4(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-((S)-3-dimethylamino-1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0681] (S)-3-(Dimethylamino)pyrrolidine gave the title compound (8 mg)

[0682] HPLC-MS (Conditions B) Retention time 1.94 min MH⁺ 546

EXAMPLE 152

[0683](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[24(S)-2-(methoxymethyl)-1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0684] (S)-2-(Methoxymethyl)pyrrolidine gave the title compound (2 mg)

[0685] HPLC-MS (Conditions B) Retention time 2.37 min MH⁺ 547

EXAMPLE 153

[0686](S)-3-[4-(3,5-Dichloro-4-2pyridylcarboxamido)1phenyl-2-2-(1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0687] Piperazine gave the title compound (1 mg)

[0688] HPLC-MS (Conditions B) Retention time 1.93 min MH⁺ 518

EXAMPLE 154

[0689](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2d(RS)-3-diethylamino-1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0690] 3-(Diethylamino)pyrrolidine gave the title compound (6 mg)

[0691] HPLC-MS (Conditions B) Retention time 1.98 min MH⁺ 574

EXAMPLE 155

[0692](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(4-(4-nitrophenyl)-1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoic

[0693] 1-(4-Nitrophenyl)piperazine gave the title compound (2 mg)

[0694] HPLC-MS (Conditions B) Retention time 2.54 min MH⁺ 639

EXAMPLE 156

[0695](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-butylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0696] Butylamine gave the title compound (3 mg)

[0697] HPLC-MS (Conditions B) Retention time 2.37 min MH⁺ 505

EXAMPLE 157

[0698](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-pentylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0699] Pentylamine gave the title compound (2 mg)

[0700] HPLC-MS (Conditions B) Retention time 2.44 min MH⁺ 519

EXAMPLE 5

[0701](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-((RS)-1-methylpropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0702] 1-Methylpropylamine gave the title compound (9 mg)

[0703] HPLC-MS (Conditions B) Retention time 2.34 min MH⁺ 505

EXAMPLE 159

[0704](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2(2-isobutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0705] Isobutylamine gave the title compound (3 mg)

[0706] HPLC-MS (Conditions B) Retention time 2 35 min MH⁺ 505

EXAMPLE 160

[0707] (S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2N-methyl-N-isopropylamino)-3,4-dioxocyclobut-1-enylamino]propanoic acid

[0708] Methylisopropylamine gave the title compound (4 mg)

[0709] HPLC-MS (Conditions B) Retention time 2.31 min MH⁺ 505

EXAMPLE 161

[0710](S)-3-4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(N-ethyl-N-methylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0711] N-Ethylmethylamine gave the title compound (6 mg)

[0712] HPLC-MS (Conditions B) Retention time 2.26 min MH⁺ 491

EXAMPLE 162

[0713] (S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[24N-methyl-N-propylamino)-3,4-dioxocyclobut-1-enylamino]propanoic acid

[0714] N-Methylpropylamine gave the title compound (3 mg)

[0715] HPLC-MS (Conditions B) Retention time 2.32 min MH⁺ 505

EXAMPLE 163

[0716](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-cyclopropanemethylamino-3,4-dioxocyclobut-1-enylamino)propanoic

[0717] Cyclopropanemethylamine gave the title compound (4 mg)

[0718] HPLC-MS (Conditions B) Retention time 2.32 min MH⁺ 503

EXAMPLE 164

[0719](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(propynylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0720] 2-Propynylamine gave the title compound (5 mg)

[0721] HPLC-MS (Conditions B) Retention time 2.26 min MH⁺ 487

EXAMPLE 165

[0722](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-isopentylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0723] Isopentylamine gave the title compound (1 mg)

[0724] HPLC-MS (Conditions B) Retention time 2.44 min MH⁺ 519

EXAMPLE 166

[0725](s)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2((RS)-2-methylbutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0726] 2-Methylbutylamine gave the title compound (2 mg)

[0727] HPLC-MS (Conditions B) Retention time 2.42 min MH⁺ 519

EXAMPLE 167

[0728](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2((RS)-1,3-dimethylbutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0729] 1,3-Dimethylbutylamine gave the title compound (3 mg)

[0730] HPLC-MS (Conditions B) Retention time 2.49 min MH⁺ 533

EXAMPLE 168

[0731](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2-(N-methyl-N-butylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0732] N-Methylbutylamine gave the title compound (3 mg)

[0733] HPLC-MS (Conditions B) Retention time 2.39 min MH⁺ 519

EXAMPLE 169

[0734](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2((RS)-1-methylbutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid 1-Methylbutylamine gave the title compound (3 mg)

[0735] HPLC-MS (Conditions B) Retention time 2.41 min MH⁺ 519

EXAMPLE 170

[0736](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-(2-allylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0737] Allylamine gave the title compound (3 mg)

[0738] HPLC-MS (Conditions B) Retention time 2.27 min MH⁺ 489

EXAMPLE 171

[0739](S)-3-[4-(3,5-Dichloro-4-1pyridylcarboxamido)phenyl]-2-[2-(2-(methylthio)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[0740] 2-(Methylthio)ethylamine gave the title compound (3 mg)

[0741] HPLC-MS (Conditions B) Retention time 2.30 min MH⁺ 523

EXAMPLE 172

[0742](S)-3-[4-(3,5-Dichloro-4-1pyridylcarboxamido)phenyl]-2-(2-carboxyethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0743] β-Alanine hydrochloride gave the title compound (4 mg)

[0744] HPLC-MS (Conditions B) Retention time 2.19 min MH⁺ 521

EXAMPLE 173

[0745](S)-3-[4-(3,5-Dichloro-4-1pyridylcarboxamido)phenyl]-2-[2((S)-1-carboxy-3-methylbutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0746] L-Leucine hydrochloride gave the title compound 0.5 mg

[0747] HPLC-MS (Conditions B) Retention time 2.35 min MH⁺ 563

EXAMPLE 174

[0748](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl-2-[2-(carboxymethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0749] Glycine hydrochloride gave the title compound (2 mg)

[0750] HPLC-MS (Conditions B) Retention time 2.19 min MH⁺ 507

EXAMPLE 175

[0751](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl-2-[2((S)-1-carboxy-2-methylpropylamino)-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0752] L-Valine hydrochloride gave the title compound (3 mg)

[0753] HPLC-MS (Conditions B) Retention time 2.28 min MH⁺ 549

EXAMPLE 176

[0754](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2((S)-1-carboxy-2-phenylethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0755] L-Phenylalanine gave the title compound (0.5 mg)

[0756] HPLC-MS (Conditions B) Retention time 2.38 min MH⁺ 597

EXAMPLE 177

[0757](S)-3-[4-3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-(2ethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0758] Ethylamine hydrochloride gave the title compound (3 mg)

[0759] HPLC-MS (Conditions B) Retention time 2.22 min MH⁺ 477

EXAMPLE 178

[0760](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-methylamino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0761] Methylamine hydrochloride gave the title compound (2 mg)

[0762] HPLC-MS (Conditions B) Retention time 2.17 MH⁺ 463

EXAMPLE 179

[0763](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-(2-dimethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0764] Dimethylamine hydrochloride gave the title compound (6 mg)

[0765] HPLC-MS (Conditions B) Retention time 2.20 min MH⁺ 477

EXAMPLE 180

[0766](S)-3-[4(3,5-Dichloro4-12pyridylcarboxamido)phenyl]-2-(2-anilino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0767] Derivatised resin (2), (320 mg) in DMF (10 ml), was treated with4-anilino-3-ethoxy-3-cyclobutene-1,2-dione (400 mg, 1.86 mmol) for 12 hat 70° then filtered and washed with DMF and DCM. The resin was treatedwith 60% trifluoroacetic acid in DCM (1.5 ml) for 3 h with agitationthen filtered. The filtrate was evaporated in vacuo to give the crudeproduct which was purified by preparative HPLC to afford the titlecompound (2 mg)

[0768] HPLC-MS (Conditions B) Retention time 2.46 min MH⁺ 525

EXAMPLE 181

[0769](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl-2-(2-phenyl-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0770] By the same method as the compound of Example 180,3-methoxy4-phenyl-3-cyclobutene-1,2-dione was used to give the titlecompound (13 mg).

[0771] HPLC-MS (Conditions B) Retention time 2.53 min MH⁺ 510

EXAMPLE 182

[0772](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-methoxy-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0773] Derivatised resin (3), (120 mg) was treated with 60%trifluoroacetic acid in DCM (1.5 ml) for 3 h with agitation thenfiltered. The filtrate was evaporated in vacuo to give the crude productwhich was purified by preparative HPLC to afford the title compound (2mg)

[0774] HPLC-MS (Conditions B) Retention time 2.26 min MH⁺ 465

EXAMPLE 183

[0775](S)-3-[4-(3,5-Dichloro-4-1pyridylcarboxamido)phenyl]-2-(2-(1-decahydroquinolyl)-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0776] Decahydroquinoline gave the title compound (1 mg).

[0777] HPLC-MS (Conditions B) Retention time 2.53 min MH⁺ 571

EXAMPLE 184

[0778] (S)-3-[4-3,5-Dichloro4-pyridylcarboxamido)1phenyl-2-[24N-benzyl-N-butylamino]-3,4-dioxocyclobut-1 enylamino]propanoic acid

[0779] N-Benzylbutylamine gave the title compound (5 mg)

[0780] HPLC-MS (Conditions B) Retention time 2.60 min MH⁺ 595

EXAMPLE 185

[0781](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(N-(2-cyanoethyl)-N-methylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0782] N-Methyl-beta-alanine nitrile gave the title compound (3 mg)

[0783] HPLC-MS (Conditions B) Retention time 2.22 min MH⁺ 516

EXAMPLE 186

[0784](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(N-(2-(2-pyridyl)ethyl)-N-methylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0785] 2-(2-Methylaminoethyl)pyridine gave the title compound (6 mg)

[0786] HPLC-MS (Conditions B) Retention time 2.03 min MH⁺ 568

EXAMPLE 187

[0787](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(1,2,3,6-tetrahydro-1-pyridyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0788] 1,2,3,6-Tetrahydropyridine gave the title compound (3 mg)

[0789] HPLC-MS (Conditions B) Retention time 2.32 min MH⁺ 515

EXAMPLE 188

[0790](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(N-methyl-N-(phenylethyl)amino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0791] N-Methylphenylethylamine gave the title compound (6 mg)

[0792] HPLC-MS (Conditions B) Retention time 2.45 min MH⁺ 567

EXAMPLE 189

[0793](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-N,N-dibutylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0794] Dibutylamine gave the title compound (5 mg)

[0795] HPLC-MS (Conditions B) Retention time 2.58 min MH⁺ 561

EXAMPLE 190

[0796](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3.3.3-trifluoropropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0797] 3,3,3-Trifluoropropylamine gave the title compound (7 mg)

[0798] HPLC-MS (Conditions B) Retention time 2.35 min MH⁺ 545

EXAMPLE 191

[0799](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(N-ethyl-N-(4pyridylmethyl)amino)-3,4-dioxocyclobut-1-enylamino]propanoic

[0800] 4-(Ethylaminomethyl)pyridine gave the title compound (4 mg)

[0801] HPLC-MS (Conditions B) Retention time 2.01 min MH⁺ 568

EXAMPLE 192

[0802](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-[2-(3-thiazolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0803] Thiazolidine gave the title compound (2 mg)

[0804] HPLC-MS (Conditions B) Retention time 2.29 min MH⁺ 521

EXAMPLE 193

[0805](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl-2-[2-(N-allyl-N-methylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0806] N-Methylallylamine gave the title compound (4 mg)

[0807] HPLC-MS (Conditions B) Retention time 2.29 min MH⁺ 503

EXAMPLE 194

[0808](S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-[2(N-benzyl-N-methylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0809] N-Benzylmethylamine gave the title compound (2 mg)

[0810] HPLC-MS (Conditions B) Retention time 2.42 min MH⁺ 553

EXAMPLE 195

[0811](S)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-N,N-iallylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0812] Diallylamine gave the title compound (5 mg)

[0813] HPLC-MS (Conditions B) Retention time 2.39 min MH⁺ 529.

EXAMPLE 196

[0814](S)-3-[4(1-Isoquinolylamino)phenyl]-2-(2-N,N-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0815] To the derivatised resin (8), (100 mg) was added ethanol (1.0 ml)and a 1M solution of diethylamine in DCM (0.7 ml). The solution wasagitated for 18 h at RT then filtered and washed thoroughly with DCM.The resin was treated with 95% trifluoroacetic acid in DCM (2.0 ml) for3 h with agitation and then filtered: The filtrate was evaporated invacuo to give the crude product which was purified by preparative HPLCto afford the title compound (2 mg).

[0816] HPLC-MS (Conditions B) Retention time 2.0 min MH⁺ 459

[0817] The following compounds of Examples 197 to 231 were prepared in asimilar manner to the compound of Example 196, each using the startingmaterial shown. For examples where the amine was added as a salt, 1 molequivalent of DIPEA was also added.

EXAMPLE 197

[0818](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-(3-methoxypropylamino)-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0819] 3-Methoxypropylamine gave the title compound (4 mg)

[0820] HPLC-MS (Conditions B) Retention time 2.0 min MH⁺ 475

EXAMPLE 198

[0821](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(1-piperidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0822] Piperidine gave the title compound (2 mg)

[0823] HPLC-MS (Conditions B) Retention time 2.1 min MH⁺ 471

EXAMPLE 199

[0824] (S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(1piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoic acid

[0825] Piperazine gave the title compound (2 mg)

[0826] HPLC-MS (Conditions B) Retention time 1.7 min MH⁺ 472

EXAMPLE 200

[0827](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-pentylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid Pentylamine gave the title compound (3 mg)

[0828] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 473

EXAMPLE 201

[0829](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0830] Propylamine gave the title compound (1 mg)

[0831] HPLC-MS (Conditions B) Retention time 2.0 min MH⁺ 445

EXAMPLE 202

[0832](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-(1-decahydroquinolinyl)-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0833] Decahydroquinoline gave the title compound (5 mg)

[0834] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 525

EXAMPLE 203

[0835](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-{2[-N-ethyl-N-(4-pyridylmethyl)amino]-3.4dioxocyclobut-1-enylamino}propanoicacid

[0836] 4-(Ethylaminomethyl)pyridine gave the title compound (2 mg)

[0837] HPLC-MS (Conditions B) Retention time 1.8 min MH⁺ 522

EXAMPLE 204

[0838](S)-3-[4(1-Isoquinolylamino)phenyl]-2-(2-tert-butylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0839] Tert-Butylamine gave the title compound (1 mg)

[0840] HPLC-MS (Conditions B) Retention time 2.1 min MH⁺ 459

EXAMPLE 205

[0841](S)-3-[4:(1-Isoquinolylamino)phenyl]-2-(2-cyclobutylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0842] Cyclobutylamine gave the title compound (1 mg)

[0843] HPLC-MS (Conditions B) Retention time 2.1 min MH⁺ 457

EXAMPLE 206

[0844](S)-3-[4(1-Isoquinolylamino)phenyl]-2-(2-thiomorpholino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0845] Thiomorpholine gave the title compound (2 mg)

[0846] HPLC-MS (Conditions B) Retention time 2.1 min MH⁺ 489

EXAMPLE 207

[0847](S)-3-[4-(1Isoquinolylamino)phenyl]-2-(2-allylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0848] Allylamine gave the title compound (0.3 mg)

[0849] HPLC-MS (Conditions B) Retention time 2.0 min MH⁺ 443

EXAMPLE 208

[0850](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(N-benzyl-N-methylamino)-3,4-dioxocyclobut-1enylamino]propanoicacid

[0851] N-Benzylmethylamine gave the title compound (5 mg)

[0852] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 507

EXAMPLE 209

[0853](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-cyclohexylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0854] Cyclohexylamine gave the title compound (3 mg)

[0855] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 485

EXAMPLE 210

[0856](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-benzylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0857] Benzylamine gave the title compound (2 mg)

[0858] HPLC-MS (Conditions B) Retention time 2.l min MH⁺ 493

EXAMPLE 211

[0859](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-{2-[3-(dimethylamino)propyl]amino-3,4-dioxocyclobut-1-enylamino}propanoicacid

[0860] 3-(Dimethylamino)propylamine gave the title compound (2 mg)

[0861] HPLC-MS (Conditions B) Retention time 1.7 min MH⁺ 488

EXAMPLE 212

[0862](S)-3-[4(1-Isoquinolylamino)phenyl]-2-[(2-pyridylmethyl)amino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0863] 2(Aminomethyl)pyridine gave the title compound (4 mg)

[0864] HPLC-MS (Conditions B) Retention time 1.9 min MH⁺ 494

EXAMPLE 213

[0865](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2(3-pyridylmethyl)amino-3,4-dioxocyclobut-1enylamino]propanoicacid

[0866] 3-(Aminomethyl)pyridine gave the title compound (1 mg)

[0867] HPLC-MS (Conditions B) Retention time 1.8 min MH⁺ 494

EXAMPLE 214

[0868](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(4-pyridylmethyl)amino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0869] 4-(Aminomethyl)pyridine gave the title compound (5 mg)

[0870] HPLC-MS (Conditions B) Retention time 1.8 min MH⁺ 494

EXAMPLE 215

[0871](s)-3-[4-1-Isoquinolylamino)phenyl]-2-[2-(2-(benzylthio)ethylamino)-3,4-dioxocyclobut-1enylamino]propanoicacid

[0872] 2-(Benzylthio)ethylamine hydrochloride gave the title compound (4mg)

[0873] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 553

EXAMPLE 216

[0874](S)-3-[4(1-Isoquinolylamino)phenyl]-2-(2-dimethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0875] Dimethylamine gave the title compound (24 mg)

[0876] HPLC-MS (Conditions B) Retention time 1.9 min MH⁺ 431

EXAMPLE 217

[0877](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-morpholino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0878] Morpholine gave the title compound (3 mg)

[0879] HPLC-MS (Conditions B) Retention time 2.0 min MH⁺ 473

EXAMPLE 218

[0880](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(N-methyl-N-butylamino)-3,4-dioxocyclobut-1enylamino]propanoicacid

[0881] N-Methylbutylamine gave the title compound (5 mg)

[0882] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 473

EXAMPLE 219

[0883](S)-3-[4(1-Isoquinolylamino)phenyl]-2-{2-[(RS)-2-methylbutylamino]-3,4-dioxocyclobut-1enylamino}propanoic acid

[0884] 2-Methylbutylamine gave the title compound (4 mg)

[0885] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 473

EXAMPLE 220

[0886] (S)-3-[4(1-Isoquinolylamino)phenyl]-2-(2-butylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0887] Butylamine gave the title compound (4 mg)

[0888] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 459

EXAMPLE 221

[0889](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-{2-[(RS)-1,3-dimethylbutylamino]-3,4-dioxocyclobut-1-enylamino}propanoicacid

[0890] 1,3-Dimethylbutylamine gave the title compound (5 mg)

[0891] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 487

EXAMPLE 222

[0892](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(N-methyl-N-isopropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[0893] Methylisopropylamine gave the title compound (3 mg)

[0894] HPLC-MS (Conditions B) Retention time 2.1 min MH⁺ 459

EXAMPLE 223

[0895](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-{2-[(RS)-1-methylbutylamino]-3,4-dioxocyclobut-1-enylamino}propanoicacid

[0896] 1-Methylbutylamine gave the title compound (6 mg)

[0897] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 473

EXAMPLE 224

[0898](S)-3-[4-(1-Isoquinolylamino)phenyl]-2-(2-isobutylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0899] Isobutylamine gave the title compound (4 mg)

[0900] HPLC-MS(Conditions B) Retention time 2.1 min MH⁺ 459

EXAMPLE 225

[0901](S)-3-[4(1-Isoquinolylamino)phenyl]-2-(2-dipropylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0902] Dipropylamine gave the title compound (4 mg)

[0903] HPLC-MS (Conditions B) Retention time 2.2 min MH⁺ 487

EXAMPLE 226

[0904](S)-3-[4(1-Isoquinolylamino)phenyl]-2-{2-[(RS)-2-methylpropylamino]-3,4-dioxocyclobut-1-enylamino}propanoicacid

[0905] 1-Methylpropylamine gave the title compound (4 mg)

[0906] HPLC-MS (Conditions B) Retention time 2.1 min MH⁺ 459

EXAMPLE 227

[0907] (S)-3-[4-(1-Isoquinolylamino)phenyl]-2-[2-(N-ethyl-N-methylamino)3,4-dioxocyclobut-1-enylamino]propanoic acid

[0908] N-Ethylmethylamine gave the title compound (1 mg)

[0909] HPLC-MS (Conditions B) Retention time 2.0 min MH⁺ 445

EXAMPLE 228

[0910](S)-3-[4-(2,3,4-Trimethoxybenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0911] To the derivatised resin (5), (120 mg) was added DCM (5 ml),DIPEA (0.1 ml, 0.6 mmol) and 2,3,4-trimethoxybenzoyl chloride (138 mg,0.6 mmol). The solution was agitated for 12 h at RT then filtered andwashed thoroughly with DCM. The resin was treated with 60%trifluoroacetic acid in DCM (1.5 ml) for 3 h with agitation and thenfiltered. The filtrate was evaporated in vacuo to give the crude productwhich was purified by preparative HPLC to afford the title compound (0.5mg).

[0912] HPLC-MS (Conditions B) Retention time 2.34 min MH⁺ 512

[0913] The following compounds of Examples 229 to 241 were prepared in asimilar manner to the compound of Example 228, each using the startingmaterial shown.

EXAMPLE 229

[0914](S)-3-[4-(2,4-Dimethoxybenzoylamino)phenyl]-2(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0915] 2,4-Dimethoxybenzoylchloride gave the title compound (2 mg)

[0916] HPLC-MS (Conditions B) Retention time 2.41 min MH⁺ 482

EXAMPLE 230

[0917](S)-3-[4-(4-Bromobenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0918] 4-Bromobenzoylchloride gave the title compound (3 mg)

[0919] HPLC-MS (Conditions B) Retention time 2.49 min MH⁺ 500

EXAMPLE 231

[0920](S)-3-[4-(2-Thienylcarbonylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0921] Thiophene-2-carbonylchloride gave the title compound (0.5 mg)

[0922] HPLC-MS (Conditions B) Retention time 2.31 min MH⁺ 428

EXAMPLE 232

[0923](S)-3-[4-(trans-Cinnamoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0924] trans-Cinnamoylchloride gave the title compound (1 mg)

[0925] HPLC-MS (Conditions B) Retention time 2.44 min MH⁺ 448

EXAMPLE 233

[0926](S)-3-[4-(Phenylacetylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0927] Phenacetylchloride gave the title compound (0.5 mg)

[0928] HPLC-MS (Conditions B) Retention time 2.34 min MH⁺ 436

EXAMPLE 234

[0929](S)-3-[4-(2,6-Dichlorobenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0930] 2,6-Dichlorobenzoylchloride gave the title compound (3 mg)

[0931] HPLC-MS (Conditions B) Retention time 2.39 min MH⁺ 490

EXAMPLE 235

[0932](S)-3-[4(2,6-Dimethylbenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0933] 2,6-Dimethylbenzoylchloride gave the title compound (1 mg)

[0934] HPLC-MS (Conditions B) Retention time 2.38 min MH⁺ 450

EXAMPLE 236

[0935](S)-3-[4-(Benzyloxyacetylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0936] Benzyloxyacetylchloride gave the title compound (1 mg)

[0937] HPLC-MS (Conditions B) Retention time 2.41 min MH⁺ 466

EXAMPLE 237

[0938](S)-3-[4-14-Cyanobenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0939] 4-Cyanobenzoylchloride gave the title compound (1 mg)

[0940] HPLC-MS (Conditions B) Retention time 2.33 min MH⁺ 447

EXAMPLE 238

[0941](S)-3-[4-(6-Chloro-3-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0942] 6-Chloronicotinylchloride gave the title compound (1 mg)

[0943] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 457

EXAMPLE 239

[0944](S)-3-[4-12-Chloro-3-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid 2-Chloronicotinylchloride gave the title compound (0.5 mg)

[0945] HPLC-MS (Conditions B) Retention time 2.18 min MH⁺ 457

EXAMPLE 240

[0946](S)-3-[4-(2-Fluorobenzoylamino)phenyl]-2-(2-proplyamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0947] 2-Fluorobenzoylchloride gave the title compound (1 mg)

[0948] HPLC-MS (Conditions B) Retention time 2.33 min MH⁺ 440

EXAMPLE 241

[0949](S)-3-[4-(3,4-Dimethoxybenzoylamino)phenyl-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0950] 3,4-Dimethoxybenzoylchloride gave the title compound (2 mg)

[0951] HPLC-MS (Conditions B) Retention time 2.28 min MH⁺ 482

EXAMPLE 242

[0952](S)-3-[4-(4-Methoxyphenoxycarbonylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0953] To the derivatised resin (5), (120 mg) was added 1,4-dioxan (4.5ml), DIPEA (0.2 ml, 1.2 mmol), water (0.5 ml) and4-methoxyphenylchloroformate (0.2 ml, 0.6 mmol). The solution wasagitated for 12 h at RT then filtered and washed thoroughly with DCM.The resin was treated with 60% trifluoroacetic acid in DCM (1.5 ml) for3 h with agitation and then filtered. The filtrate was evaporated invacuo to give the crude product which was purified by preparative HPLCto afford the title compound (2 mg).

[0954] HPLC-MS (Conditions B) Retention time 2.42 min MH⁺ 468

[0955] The following compounds of Examples 243 to 246 were prepared in asimilar manner to the compound of Example 242, each using the startingmaterial shown.

EXAMPLE 243

[0956](S)-3-[4-(4-Methylphenoxycarbonylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0957] p-Tolylchloroformate gave the title compound (0.5 mg)

[0958] HPLC-MS (Conditions B) Retention time 2.50 min MH⁺ 452

EXAMPLE 244

[0959](S)-3-[4-(4-Fluorophenoxycarbonylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-t-enylamino)propanoicacid 4-Fluorophenylchloroformate gave the title compound (2 mg)

[0960] HPLC-MS (Conditions B) Retention time 2.45 min MH⁺ 456

EXAMPLE 245

[0961] (S)-3-[44Phenoxycarbonylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0962] Phenylchloroformate gave the title compound (2 mg)

[0963] HPLC-MS (Conditions B) Retention time 2.42 min MH⁺ 438

EXAMPLE 246

[0964](S)-3-[4-(4-Nitrobenzyloxycarbonylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0965] 4-Nitrobenzylchloroformate gave the title compound (1 mg)

[0966] HPLC-MS (Conditions B) Retention time 2.47 min MH⁺ 497

EXAMPLE 247

[0967](S)-3-(4-3enzoylphenyl)-2-(2-propylamino-3.4dioxocyclobut-1-enylamino)propanoicacid

[0968] To the derivatised resin (9), (200 mg) was added ethanol (1.6ml), DCM (0.4 ml) and propylamine (0.08 ml, 1 mmol). The solution wasagitated for 12 h at RT then filtered and washed thoroughly with DCM.The resin was treated with 95% trifluoroacetic acid in DCM (2.0 ml) for3 h with agitation and then filtered. The filtrate was evaporated invacuo to give the crude product which was purified by preparative HPLCto afford the title compound (4 mg).

[0969] HPLC-MS (Conditions B) Retention time 2.4 min MH⁺ 407.

[0970] The following compound of Example 248 was prepared in a similarmanner to the compound of Example 247 using the starting material shown.

EXAMPLE 248

[0971](S)-3-(4-Benzoylphenyl)-2-(2-morpholino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0972] Morpholine gave the title compound (5 mg)

[0973] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 435

EXAMPLE 249

[0974](S)-3-[4-(1-Isoquinolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0975] A slurry of derivatised resin (5) (prepared from Wang resin (0.7mmol/g), 100 mg) in DCM (5 ml) was treated with 1-isoquinolinecarboxylic acid (56 mg, 0.30 mmol), DIEA (45 μl, 0.25 mmol) and[O-(7-azabenzotriazol-1-yl)-1,1,3,3-Tetramethyluronium-hexafluorophosphate](HATU) (95 mg, 0.25 mmol). The mixture was agitated for 16 h at RT thenfiltered and washed thoroughly with DCM, DMF, MeOH, DMF then DCM. Theresin was treated with 50% trifluoroacetic acid in DCM (5 ml) for 3 hwith agitation and then filtered. The resin was washed with a furtherportion of DCM (5 ml). The combined filtrate was evaporated in vacuo togive the crude product which was purified by preparative HPLC to affordthe title compound (7.3 mg).

[0976] HPLC-MS (Conditions B). Retention time 2.47 min, MH⁺ 473 Thefollowing compounds of Examples 250 to 281 were prepared in a similarmanner to the compound of Example 249, each using the starting materialshown.

EXAMPLE 250

[0977](S)-3-}4-[2-Benzo(b)furanylcarboxamido]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0978] 2-Benzo(b)furancarboxylic acid gave the title compound (1.0 mg)

[0979] HPLC-MS (Conditions B). Retention time 2.45 min, MH⁺ 462

EXAMPLE 251

[0980](S)-3-[4-(4-Methoxy-2-quinolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0981] 4-Methoxy-2-quinolinecarboxylic acid gave the title compound (5.3mg)

[0982] HPLC-MS (Conditions B). Retention time 2.59 min, MH⁺ 503

EXAMPLE 252

[0983](S)-3-{4-[4-(4-Oxo4,5,6,7,-tetrahydrobenzo(b)furan-3-ylcarboxamido]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0984] 4-Oxo-4,5,6,7-tetrahydrobenzo(b)furan-3-carboxylic acid gave thetitle compound (8.2 mg)

[0985] HPLC-MS (Conditions B). Retention time 2.37 min, MH⁺ 480

EXAMPLE 253

[0986](S)-3-[4-(2-(1-Pyrrolyl)-5-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0987] 2-(1-Pyrrolyl)-5-pyridinecarboxylic acid gave the title compound(1.7 mg)

[0988] HPLC-MS (Conditions B). Retention time 2.45 min, MH⁺ 488

EXAMPLE 254

[0989](S)-3-[4-(3-Indazolylcarboamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0990] 3-Indazolecarboxylic acid gave the title compound (5.0 mg)

[0991] HPLC-MS (Conditions B). Retention time 2.34 min, MH⁺ 462

EXAMPLE 255

[0992](S)-3-[4-(4-Fluorobenzoylamino)phenyl]-22-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0993] 4-Fluorobenzoic acid gave the title compound (3.7 mg)

[0994] HPLC-MS (Conditions B). Retention time 2.37 min, MH⁺ 440

EXAMPLE 256

[0995](S)-3-[4-(4-Methoxybenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0996] 4-Methoxybenzoic acid gave the title compound (0.3 mg)

[0997] HPLC-MS (Conditions B). Retention time 2.34 min, MH⁺ 452

EXAMPLE 257

[0998](S)-3-[4-(4-Acetamidobenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[0999] 4-Acetamidobenzoic acid gave the title compound (3.7 mg)

[1000] HPLC-MS (Conditions B). Retention time 2.16 min, MH⁺ 479

EXAMPLE 258

[1001](S)-3-[4-(4-Acetylbenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1002] 4-Acetylbenzoic acid gave the title compound (2.0 mg)

[1003] HPLC-MS (Conditions B). Retention time 2.28 min, MH⁺ 461

EXAMPLE 259

[1004](S)-3-[4-(4-Nitrobenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1005] 4-Nitrobenzoic acid gave the title compound (4.3 mg)

[1006] HPLC-MS (Conditions B). Retention time 2.39 min, MH⁺ 467

EXAMPLE 260

[1007](S)-3-{4-[4-(4-Hydroxyphenyl)benzoylamino]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylaminolpropanoicacid

[1008] 4-Hydroxybiphenyl carboxylic acid gave the title compound (0.8mg)

[1009] HPLC-MS (Conditions B). Retention time 2.36 min, MH⁺ 514

EXAMPLE 261

[1010](S)-3-[4-(4-Cyanobenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1011] 4-Cyanobenzoic acid gave the title compound (6.5 mg)

[1012] HPLC-MS (Conditions B). Retention time 2.32 min, MH⁺ 447

EXAMPLE 262

[1013](S)-3-[4-(4-Trifluoromethylbenzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1014] 4-Trifluoromethylbenzoic acid gave the title compound (5.4 mg)

[1015] HPLC-MS (Conditions B). Retention time 2.55 min, MH⁺ 560

EXAMPLE 263

[1016](S)-3-[4-(N-Oxo4-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1017] 4-Pyridyl-N-oxide carboxylic acid gave the title compound (4.7mg)

[1018] HPLC-MS (Conditions B). Retention time 1.97 min, MH⁺ 439

EXAMPLE 264

[1019](S)-3-[4-(2,6-Dichloro-3-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1020] 2,6, Dichloronicotinic acid gave the title compound (4.7 mg)

[1021] HPLC-MS (Conditions B). Retention time 2.31 min, MH⁺ 493

EXAMPLE 265

[1022](S)-3-[4-(2-(Methoxycarbonyl)benzoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1023] 2-Methoxycarbonylbenzoic acid gave the title compound (3.4 mg)

[1024] HPLC-MS (Conditions B). Retention time 2.28 min, MH⁺ 480

EXAMPLE 266

[1025](S)-3-{4-[5-Methyl-2-(trifluoromethyl)-3furanylcarboxamido]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1026] 5-Methyl-2-(trifluoromethyl)-3-furancarboxylic acid gave thetitle compound (5.6 mg)

[1027] HPLC-MS (Conditions B). Retention time 2.48 min, MH⁺ 494

EXAMPLE 267

[1028](S)-3-[4-(2-Acetyl-3-thienylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1029] 2-Acetyl-3-thiophenecarboxylic acid gave the title compound (5.2mg)

[1030] HPLC-MS (Conditions B). Retention time 2.28 min, MH⁺ 470

EXAMPLE 268

[1031](S)-3-{4-[((R)-2-Oxothiazolidin-4-ylcarboxamido]phenyl}-2-[2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1032] (R)-2-Oxothiazolidine-4-carboxylic acid gave the title compound(5.4 mg)

[1033] HPLC-MS (Conditions B). Retention time 2.07 min, MH⁺ 447

EXAMPLE 269

[1034](S)-3-[4-(4-Nitro-3-pyrazolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid 4-Nitro-3-pyrazolecarboxylic acid gave the title compound (3.0 mg)

[1035] HPLC-MS (Conditions B). Retention time 2.14 min, MH⁺ 457

EXAMPLE 270

[1036](S)-3-[4-(5-Chloro-2-thienylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1037] 5-Chloro-2-thiophenecarboxylic acid gave the title compound (5.3mg)

[1038] HPLC-MS (Conditions B). Retention time 2.48 min, MH⁺ 462

EXAMPLE 271

[1039] (S)-3-[4-(1-Methyl-5-nitro-4-pyrazolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoic acid

[1040] 1-Methyl-5-nitro-4-pyrazolecarboxylic acid gave the titlecompound (6.1 mg)

[1041] HPLC-MS (Conditions B). Retention time 2.23 min, MH⁺ 471

EXAMPLE 272

[1042](S)-3-[4-(2-Furoylamino)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1043] 2-Furoic acid gave the title compound (3.7 mg)

[1044] HPLC-MS (Conditions B). Retention time 2.23 min, MH⁺ 412

EXAMPLE 273

[1045](S)-3-[4-(2,4-Dimethyl-5-thiazolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1046] 2,4-Dimethyl-5-thiazolecarboxylic acid gave the title compound(4.2 mg)

[1047] HPLC-MS (Conditions B). Retention time 2.18 min, MH⁺ 457

EXAMPLE 274

[1048](S)-3-[4-(1,2,3-thiadiazol-4-ylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1049] 1,2,3,Thiadiazole-5-carboxylic acid gave the title compound (4.9mg)

[1050] HPLC-MS (Conditions B). Retention time 2.20 min, MH⁺ 430

EXAMPLE 275

[1051](S)-3-[4-(2-Thienylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1052] 2-Thiophenecarboxylic acid gave the title compound (5.0 mg)

[1053] HPLC-MS (Conditions B). Retention time 2.31 min, MH⁺ 428

EXAMPLE 276

[1054](S)-3-[4-(2-Pyrazinylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-t-enylamino)propanoicacid

[1055] 2-Pyrazinecarboxylic acid gave the title compound (4.2 mg)

[1056] HPLC-MS (Conditions B). Retention time 2.16 min, MH⁺ 424

EXAMPLE 277

[1057](S)-3-{4-[(2-Furyl)oxalylamino]phenyl}-2-(2-propylamino-3,4-dioxocyclobut-1-enylamio)propanoicacid

[1058] α-Oxo-2-furanacetic acid gave the title compound (4.8 mg)

[1059] HPLC-MS (Conditions B). Retention time 2.3 min, MH⁺ 440.

EXAMPLE 278

[1060](S)-3-[4-(3-Methyl-2-thienylcarboxamido)phenyl]-2-(2-2propylamino-3,4-dioxocyclobut-1-enylaminolpropanoicacid

[1061] 3-Methyl-2-thiophenecarboxylic acid gave the title compound (2.0mg)

[1062] HPLC-MS (Conditions B). Retention time 2.37 min, MH⁺ 442

EXAMPLE 279

[1063](S)-3-[4-(4-Methyl-1,2,3-thiadiazol-5-ylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1064] 4-Methyl-1,2,3-thiazole-5carboxylic acid gave the title compound(4.0 mg)

[1065] HPLC-MS (Conditions B). Retention time 2.24 min, MH⁺ 444

EXAMPLE 280

[1066](S)-3-[4-(5-Phenyl-4-oxazolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1067] 5-Phenyl-4-oxazolecarboxylic acid gave the title compound (5.9mg)

[1068] HPLC-MS (Conditions B). Retention time 2.51 min, MH⁺ 489

EXAMPLE 281

[1069](S)-3-[4-(3-Methyl-5-trifluoromethyl-4-isoxazolylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoic acid

[1070] 3-Methyl-5-trifluoromethyl-4-isoxazolecarboxylic acid gave thetitle compound (5.8 mg)

[1071] HPLC-MS (Conditions B). Retention time 2.43 min, MH⁺ 495.

[1072] The following compounds of Examples 282 to 323 were prepared in asimilar manner to the compound of Example 105, using derivatised resin(4) and the starting material shown. For examples where the amine wasadded as a salt 1 mol equivalent of DIPEA was also added.

EXAMPLE 282

[1073](RS)-3-[4-(3,5-Dichloro-4-pyrodylcarboxamido)phenyl]-3-[2-(2-morpholinoethylamino)-3,4-dioxocyclobut-1enylamino]propanoic acid

[1074] N-(2-Aminoethyl)morpholine gave the title compound (2 mg)

[1075] HPLC-MS (Conditions B) Retention time 1.98 min MH⁺ 562

EXAMPLE 283

[1076](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(2-piperdinoethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1077] 1-(2-Aminoethyl)piperidine gave the title compound (2mg)

[1078] HPLC-MS (Conditions B) Retention time 2.02 min MH⁺ 560

EXAMPLE 284

[1079](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3-(2-oxopyrrolidin-1-yl)propylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1080] 1-(3-Aminopropyl)-2-pyrrolidinone gave the title compound (1 mg)

[1081] HPLC-MS (Conditions B) Retention time 2.14 min MH⁺ 574

EXAMPLE 285

[1082](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)1phenyl-3-[2-(3-(1-imidazolyl)propylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[1083] N-(3-Aminopropyl)imidazole gave the title compound (3 mg)

[1084] HPLC-MS (Conditions B) Retention time 1.98 min MH⁺ 557

EXAMPLE 286

[1085](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl-3-[2-(1-benzyl-4-piperidinylamino]-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1086] 4-Amino-1-benzylpiperidine gave the title compound (6 mg)

[1087] HPLC-MS (Conditions B) Retention time 2.13 min MH⁺ 622

EXAMPLE 287

[1088](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(2-pyridylmethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1089] 2-(Aminomethyl)pyridine gave the title compound (6 mg)

[1090] HPLC-MS (Conditions B) Retention time 2.17 min MH⁺ 540

EXAMPLE 288

[1091](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3-pyridylmethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1092] 3-(Aminomethyl)pyridine gave the title compound (3 mg)

[1093] HPLC-MS (Conditions B) Retention time 2.07 min MH⁺ 540

EXAMPLE 289

[1094](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3,3-dimethylbutylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1095] 3,3-Dimethylbutylamine gave the title compound (3 mg)

[1096] HPLC-MS (Conditions B) Retention time 2.45 min MH⁺ 533

EXAMPLE 290

[1097](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3,4-dichlorobenzylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1098] 3,4-Dichlorobenzylamine gave the title compound (4 mg)

[1099] HPLC-MS (Conditions B) Retention time 2.51 min MH⁺ 607

EXAMPLE 291

[1100](RS)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-3-[2-(2-(1-piperazinyl)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[1101] N-(2-Aminoethyl)piperazine gave the title compound (1 mg)

[1102] HPLC-MS (Conditions B) Retention time 1.97 min MH⁺ 561

EXAMPLE 292

[1103](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-isopropylamino-3,4-dioxocyclobut-1-enylamino)1propanoicacid

[1104] Isopropylamine gave the title compound (1 mg)

[1105] HPLC-MS (Conditions B) Retention time 2.25 min MH⁺ 491

EXAMPLE 293

[1106](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-propylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1107] Propylamine gave the title compound (2 mg)

[1108] HPLC-MS (Conditions B) Retention time 2.25 min MH⁺ 491

EXAMPLE 294

[1109](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-tert-butylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1110] Tert-Butylamine gave the title compound (0.5 mg)

[1111] HPLC-MS (Conditions B) Retention time 2.33 min MH⁺ 505

EXAMPLE 295

[1112](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-benzylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1113] Benzylamine gave the title compound (1 mg)

[1114] HPLC-MS (Conditions B) Retention time 2.37 min MH⁺ 539

EXAMPLE 296

[1115](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3-(dimethylaminoloropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1116] 3-(Dimethylamino)propylamine gave the title compound (0.5 mg)

[1117] HPLC-MS (Conditions B) Retention time 1.89 min MH⁺ 534

EXAMPLE 297

[1118](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3-isopropoxypropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1119] 3-Isopropoxypropylamine gave the title compound (1 mg)

[1120] HPLC-MS (Conditions B) Retention time 2.3 min MH⁺ 549

EXAMPLE 298

[1121](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3-ethoxypropylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1122] 3-Ethoxypropylamine gave the title compound (2 mg)

[1123] HPLC-MS (Conditions B) Retention time 2.23 min MH⁺ 535

EXAMPLE 299

[1124]IRS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(2methoxyethylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1125] 2-Methoxyethylamine gave the title compound (0.5 mg)

[1126] HPLC-MS (Conditions B) Retention time 2.16 min MH⁺ 507

EXAMPLE 300

[1127](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(3-methoxyoropylamino)-3,4-dioxocyclobut-1enylaminolpropanoicacid

[1128] 3-Methoxypropylamine gave the title compound (0.5 mg)

[1129] HPLC-MS (Conditions B) Retention time 2.18 min MH⁺ 521

EXAMPLE 301

[1130](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-cyclobutylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1131] Cyclobutylamine gave the title compound (2 mg)

[1132] HPLC-MS (Conditions B) Retention time 2.28 min MH⁺ 503

EXAMPLE 302

[1133](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-cyclopropylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1134] Cyclopropylamine gave the title compound (2 mg)

[1135] HPLC-MS (Conditions B) Retention time 2.19 min MH⁺ 489

EXAMPLE 303

[1136](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-2-(2-(benzylthio)ethylamino)-3,4-dioxocyclobut-1-enylamino]propanoic

[1137] 2-(Benzylthio)ethylamine hydrochloride gave the title compound(0.5 mg)

[1138] HPLC-MS (Conditions B) Retention time 2.46 min MH⁺ 599

EXAMPLE 304

[1139] (RS)-3-[4(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(4-(1,2,3-thiadiazol4-yl)benzylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1140] 4-(1,2,3-Thiadiazol4-yl)benzylamine gave the title compound (2mg)

[1141] HPLC-MS (Conditions B) Retention time 2.38 min MH⁺ 623

EXAMPLE 305

[1142](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-cyclohexylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1143] Cyclohexylamine gave the title compound (0.5 mg)

[1144] HPLC-MS (Conditions B) Retention time 2.39 min MH⁺ 531

EXAMPLE 306

[1145](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-piperidinyl-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1146] Piperidine gave the title compound (2 mg)

[1147] HPLC-MS (Conditions A) Retention time 2.32 min MH⁺ 517

EXAMPLE 307

[1148](RS)-3-[4-(3,5-Dichloro-4-2pyridylcarboxamido)phenyl]-3-(2-thiomorpholino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1149] Thiomorpholine gave the title compound (1 mg)

[1150] HPLC-MS (Conditions A) Retention time 2.32 min MH⁺ 535

EXAMPLE 308

[1151](RS)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-3-[2-(4-methyl-1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1152] 1-Methylpiperazine gave the title compound (3 mg)

[1153] HPLC-MS (Conditions A) Retention time 1.93 min MH⁺ 532

EXAMPLE 309

[1154](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-diethylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1155] Diethylamine gave the title compound (2 mg)

[1156] HPLC-MS (Conditions A) Retention time 2.29 min MH⁺ 505

EXAMPLE 310

[1157](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1158] Pyrrolidine gave the title compound (1 mg)

[1159] HPLC-MS (Conditions A) Retention time 2.24 min MH⁺ 503

EXAMPLE 311

[1160](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(4-ethyl-1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1161] 1-Ethylpiperazine gave the title compound (1 mg)

[1162] HPLC-MS (Conditions A) Retention time 1.94 min MH⁺ 546

EXAMPLE 312

[1163](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(4-(hydroxyoropyl)-1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1164] 1-Piperazinepropanol gave the title compound (3 mg)

[1165] HPLC-MS (Conditions A) Retention time 1.93 min MH⁺ 576

EXAMPLE 313

[1166](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(1-piperazinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1167] Piperazine gave the title compound (4 mg)

[1168] HPLC-MS (Conditions A) Retention time 1.92 min MH⁺ 518

EXAMPLE 314

[1169] (RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-((S)3-dimethylamino-1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1170] (S)-3-(Dimethylamino)pyrrolidine gave the title compound (3 mg)

[1171] HPLC-MS (Conditions A) Retention time 1.92 min MH⁺ 546

EXAMPLE 315

[1172](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-((RS)-3-diethylamino-1-pyrrolidinyl)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1173] 3-(Diethylamino)pyrrolidine gave the title compound (3 mg)

[1174] HPLC-MS (Conditions A) Retention time 1.95 min MH⁺ 574

EXAMPLE 316

[1175](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-butylamino-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1176] Butylamine gave the title compound (0.1 mg)

[1177] HPLC-MS (Conditions A) Retention time 2.33 min MH⁺ 505

EXAMPLE 317

[1178](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-pentylamino-3,4-dioxocyclobut-1enylamino)propanoicacid

[1179] Pentylamine gave the title compound (1 mg)

[1180] HPLC-MS (Conditions A) Retention time 2.40 min MH⁺ 519

EXAMPLE 3198

[1181](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-((RS)-2-butylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1182] 1-Methylpropylamine gave the title compound (2 mg)

[1183] HPLC-MS (Conditions A) Retention time 2.31 min MH⁺ 505

EXAMPLE 319

[1184](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-(2-isobutylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1185] Isobutylamine gave the title compound (2 mg)

[1186] HPLC-MS (Conditions A) Retention time 2.31 min MH⁺ 505

EXAMPLE 320

[1187](RS)-3-[4(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(N-methyl-N-isopropylamino)-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1188] Methylisopropylamine gave the title compound (2 mg)

[1189] HPLC-MS (Conditions A) Retention time 2.3 min MH⁺ 505

EXAMPLE 321

[1190](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(N-ethyl-N-methylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1191] N-Ethylmethylamine gave the title compound (0.2 mg)

[1192] HPLC-MS (Conditions A) Retention time 2.24 min MH⁺ 491

EXAMPLE 322

[1193](RS)-3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(N-methyl-N-butylamino)-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1194] N-Methylbutylamine gave the title compound (0.3 mg)

[1195] HPLC-MS (Conditions A) Retention time 2.38 min MH⁺ 519

EXAMPLE 323

[1196](RS)-3-[4(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-3-[2-(N-ethyl-N-(pyridylmethyl)amino]-3,4-dioxocyclobut-1-enylamino]propanoicacid

[1197] 4-(Ethylaminomethyl)pyridine gave the title compound (1 mg)

[1198] HPLC-MS (Conditions A) Retention time 2.01 min MH⁺ 568

EXAMPLE 324

[1199]3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl-2-(2-isopropylamino-3,4-dioxocyclobut-1-enylamino)propanoicacid

[1200] Derivatised resin 6 (1.0 g) was treated with the Intermediate 44(0.4 g, 3.0 mmol) and a catalytic amount of dirhodiumtetraacetate intoluene (10 ml) at 1200 for 2.5 h. The resin was then filtered andwashed with DMF and DCM to give resin boundα-(2-methoxy-3,4-dioxocyclo-but-1-enylamino)diethylphosphonoacetate.This resin was then treated with4-(3,5-dichloro-4-pyridylcarboxamido)benzaidehyde (0.53 g, 1.5 mmol) anddiazabicycloundec-7-ene (DBU) (0.1 g, 1.2 mmol) in DCM (5.0 ml). Themixture was agitated at ambient temperature for 72 h then filtered andthe resin washed thoroughly with DCM. A 90 mg portion of this resin wastreated with 2-propylamine (0.045 mL, 0.6 mmol), in DCM (0.2 mL) andMeOH (0.8 mL). The mixture was agitated at ambient temperature for 16 hthen filtered and washed thoroughly with DCM, MeOH, DMF, MeOH and DCM.The resin was treated with 50% trifluoroacetic acid in DCM (2 ml) for 3h with agitation and then filtered. The resin was washed with a furtherportion of DCM (2 ml) and the combined filtrate was evaporated in vacuoto give the crude product which was purified by preparative HPLC toafford the title compound (0.9 mg).

[1201] HPLC-MS (Conditions B). Retention time 2.29 min, MH⁺ 489 Thefollowing compound of Example 325 was prepared in an identical manner tothe compound of Example 324, using the starting material shown.

EXAMPLE 325

[1202]3-[4-(3,5-Dichloro-4-pyridylcarboxamido)phenyl]-2-(2-cyclobutylamino-3,4-dioxocyclobut-1-enylamino)prop-2-enoicacid

[1203] Cyclobutylamine gave the title compound (0.4 mg)

[1204] HPLC-MS (Conditions B). Retention time 2.33 min, MH⁺ 501.

[1205] The following assays can be used to demonstrate the potency andselectivity of the compounds according to the invention. In each ofthese assays an IC₅₀ value was determined for each test compound andrepresents the concentration of compound necessary to achieve 50%inhibition of cell adhesion where 100%=adhesion assessed in the absenceof the test compound and 0%=absorbance in wells that did not receivecells.

[1206] α₄β₁ Integrin-dependent Jurkat cell adhesion to VCAM-Ig

[1207] 96 well NUNC plates were coated with F(ab)₂ fragment goatanti-human IgG Fcγ-specific antibody [Jackson Immuno Research109-006-098: 100 μl at 2 μg/ml in 0.1M NaHCO₃, pH 8.4], overnight at 40.The plates were washed (3×) in phosphate-buffered saline (PBS) and thenblocked for 1 h in PBS/1% BSA at RT on a rocking platform. After washing(3×in PBS) 9 ng/ml of purified 2d VCAM-Ig diluted in PBS/1% BSA wasadded and the plates left for 60 minutes at RT on a rocking platform.The plates were washed (3×in PBS) and the assay then performed at 370for 30 min in a total volume of 200 μl containing 2.5×10⁵ Jurkat cellsin the presence or absence of titrated test compounds.

[1208] Each plate was washed (2×) with medium and the adherent cellswere fixed with 100 μl MeOH for 10 minutes followed by another wash. 100μl 0.25% Rose Bengal (Sigma R4507) in PBS was added for 5 minutes at RTand the plates washed (3×) in PBS. 100 μl 50% (v/v) ethanol in PBS wasadded and the plates left for 60 min after which the absorbance (570 nm)was measured.

[1209] α₄β₇Integrin-dependent JY cell adhesion to MAdCAM-Ig

[1210] This assay was performed in the same manner as the α₄β₁ assayexcept that MAdCAM-Ig (150 ng/ml) was used in place of 2d VCAM-Ig and asub-line of the β-lympho blastoid cell-line JY was used in place ofJurkat cells. The IC₅₀ value for each test compound was determined asdescribed in the α₄β₁ integrin assay.

[1211] α5β₁ Integrin-dependent K562 cell adhesion to fibronectin

[1212] 96 well tissue culture plates were coated with human plasmafibronectin (Sigma F0895) at 5 μg/ml in phosphate-buffered saline (PBS)for 2 hr at 37° C. The plates were washed (3×in PBS) and then blockedfor 1 h in 100 μl PBS/1% BSA at RT on a rocking platform. The blockedplates were washed (3×in PBS) and the assay then performed at 37° C. ina total volume of 200 μl containing 2.5×10⁵ K562 cells,phorbol-12-myristate-13-acetate at 10 ng/ml, and in the presence orabsence of titrated test compounds. Incubation time was 30 minutes. Eachplate was fixed and stained as described in the α₄β₁ assay above

[1213] α_(m)β₂-dependent human polymorphonuclear neutrophils adhesion toplastic

[1214] 96 well tissue culture plates were coated with RPMI 1640/10% FCSfor 2 h at 37° C. 2×10⁵ freshly isolated human venous polymorphonuclearneutrophils (PMN) were added to the wells in a total volume of 200 μl inthe presence of 10 ng/ml phorbol-12-myristate-13-acetate, and in thepresence or absence of test compounds, and incubated for 20 min at 37°C. followed by 30 min at RT. The plates were washed in medium and 100 μl0.1% (w/v) HMB (hexadecyl trimethyl ammonium bromide, Sigma H5882) in0.05M potassium phosphate buffer, pH 6.0 added to each well. The plateswere then left on a rocker at RT for 60 min. Endogenous peroxidaseactivity was then assessed using tetramethyl benzidine (TMB) as follows:PMN lysate samples mixed with 0.22% H₂O₂ (Sigma) and 50 μg/ml TMB(Boehringer Mannheim) in 0.1M sodium acetate/citrate buffer, pH 6.0 andabsorbance measured at 630 nm.

[1215] αllb/β₃-dependent human platelet aggregation

[1216] Human platelet aggregation was assessed using impedanceaggregation on the Chronolog Whole Blood Lumiaggregometer. Humanplatelet-rich plasma (PRP) was obtained by spinning fresh human venousblood anticoagulated with 0.38% (v/v) tri-sodium citrate at 220×g for 10min and diluted to a cell density of 6×10⁸/ml in autologous plasma.Cuvettes contained equal volumes of PRP and filtered Tyrode's buffer(g/liter: NaCl 8.0; MgCl₂.H₂O 0.427; CaCl₂ 0.2; KCl 0.2; D-glucose 1.0;NaHCO₃ 1.0; NaHPO₄.2H₂O 0.065). Aggregation was monitored followingaddition of 2.5CM ADP (Sigma) in the presence or absence of inhibitors.

[1217] In the above assays the preferred compounds of the invention inwhich R¹ is an α₄ integrin binding group, such as the compounds of theExamples generally have IC₅₀ values in the α₄β₁ and α₄ β₇ assays of 1 μMand below. In the other assays featuring at integrins of other subgroupsthe same compounds had IC₅₀ values of 50 μM and above thus demonstratingthe potency and selectivity of their action against α₄ integrins.

[1218] The following assays may be used to determine the ability ofcompounds according to the invention to inhibit α_(v)β₃ and α_(v)β₅function.

[1219] α_(v)β₃-Dependent Direct Binding Assay

[1220] 96 Well NUNC immunoplates were coated overnight with anon-blocking anti-β₃ monoclonal antibody at 2 μg/ml in Dulbecco'sphosphate buffered saline (PBS) and subsequently blocked with 5%9W/v)BSA in PBS (Sigma, fraction V) for 60 min. at RT. After washing inTris-buffered saline (TBS: 20 mM Tris/150 mM NaCl, pH 7.5), plates thenreceived 100 μl of a lysate prepared from JY cells and were incubatedfor 3 h at RT. The lysate was made by lysing JY B-lymphoblastoid cellsat 5×10⁷ cells were ml in TBS containing 1 mM MnCl₂, 1% (V/v) BSA/0.1%(v/v) Tween 20 and were incubated for a further 2 hours at RT.Inhibitors were titrated into the fibronectin prior to addition toplates. After washing, streptavidin-peroxidase (Amersham) at 1:500 inTBS/1% (W/v) BSA/0.1% (v/v)Tween 20 was added and plates incubated for 1h at RT. Finally 100 μl TMB substrate was added and Absorbance (630 nm)measured after 10-15 miunbutes. IC₅₀ values for inhibition of adhesionwere calculated on the Activity Base curve fitting programme.

[1221] α_(v)β₃-Dependent Cell Adhesion Assay

[1222] This was a modification of a published method [Stupack et al.,Exp. Cell. Tes. 203, 443-448 (1992)] and employed the JY cell line.These cells are maintained in RPMI 1640+10% FCS+2 mM L-glutamine and,when used for assay, were washed in assat medium (RPMI 1640+10% FCS),suspended at 4×10⁶/ml in the same medium and pretreated with a blockingmonoclonal antibody to CD18 (6.5E, F(ab′)₂ fragment) for 10 min at RT.96 Well NUNC immunoplates were coated with 100 μl 2.5uk/μl humanvitronectin in PBS per well for 2 h at 37° C.; they were then washed2×in PBS and blocked with 1% (w/v) BSA in PBS for 60 min at RT andwashed 2×more in PBS. 2×1-5 JY per well were added to wells containingcompounds serially titrated across the plate and, finally,phorbol-12-myristate-13-acetate at 10 ng/ml was added in a final volumeof 200 μl. After incubation at 370C for 30 min, non-adherent cells wereremoved by washing 3×in assay medium, adherent cells were fixed in MeOHand stained with 0.25% (W/v) Rose Bengal in PBS for 5 min, unbound dyewas removed by 3 further washes in PBS and cell-bound dye was releasedwith 1:1 PBS:ethanol. Absorbance at 570 nm was then measured. IC₅₀values for inhibition of adhesion were calculated as described above forthe direct binding assay.

[1223] α_(v)β₅-Dependent Cell Adhesion Assay

[1224] This assay was based on a published method [Koivunen et al, J.Bio. Chem. 268, 20205-20210 (1993)] and employed the human colonadenocarcinoma cell line HT-29. HT-29 Cells were routinely maintained inDMEM+10% FCS+2 mM L-glutamine and were removed from flasks usingtrypsin/EDTA, washed 2×in assay medium and suspended at 4×10⁶/ml in thesame medium. The cells were allowed to ‘rest’ for 15 min. at RT beforebeing added (2×10⁵/well) to wells containing compounds serially titratedacross the plate in a final volume of 200 μl. The 96 well NUNCimmunoplates had been coated with human vit ronectin as described abovefor the α_(v)β₃ assay. After incubation at 37° C. for 60 min, adhesionwas assessed as described above for the α_(v)β₃ assay.

[1225] In the above assays the preferred compounds of the inventiongenerally have IC₅₀ values of 1 μM and below.

[1226] The advantageous clearance properties of compounds according tothe invention may be demonstrated as follows:

[1227] Hepatic clearance, whether metabolic or biliary, can make asubstantial contribution to the total plasma clearance of a drug. Thetotal plasma clearance is a principal parameter of the pharmacokineticproperties of a medicine. It has a direct impact on the dose required toachieve effective plama concentrations and has a major impact on theelimination half-life and therefore the dose-interval. Furthermore, highhepatic clearance is 3n indicator of high first-pass hepatic clearanceafter oral administration and therefore low oral bioavailability.

[1228] Many peptidic and non-peptidic carboxylic acids of therapeuticinterest are subject to high hepatic clearance from plasma. Except fordrugs which function in the liver, hepatic uptake from blood or plasmais undesirable because it leads to high hepatic clearance if thecompound is excreted in bile or metabolised, or if the substance is notcleared from the liver, it may accumulate in the liver and interferewith the normal function of the liver.

[1229] The total plasma clearance of a compound according to theinvention can be determined as follows:

[1230] a small dose of the compound in solution is injected into a veinof a test animal. Blood samples are withdrawn from a blood vessel of theanimal at several times after the injection, and the concentration ofcompound in the bleed or plasma is measured using a suitable assay. Thearea under the curve (AUCiv) is calculated by non-compartmental methods(for example, the trapezium method) or by pharmacokinetic modelling. Thetotal plasma clearance (CLp) is calculated by dividing the intravenousdose(Div) by the AUCiv for the blood plasma concentration—time course ofa drug administered by the intravenous route: CLp =Div÷AUCiv

[1231] When tested in this manner, compounds according to the inventionare not rapidly or extensively extracted by the liver and have low totalplasma clearance where low is defined as less than 10 ml/min/kg in thelaboratory rat (Sprague Dawley CD). This compares favourably withfunctionally equivalent integrin binding compounds in which the squaricacid framework of compounds of formula (1) is not present.

1. A compound of formula (1):

wherein R¹ is an integrin binding group; R² is a hydrogen atom or aC₁₋₆alkyl group; L¹ is a covalent bond or a linker atom or group; n iszero or the integer 1; Alk¹ is an optionally substituted aliphaticchain; R³ is a hydrogen atom or an optionally substitutedheteroaliphatic, cycloaliphatic, heterocycloaliphatic,polycycloaliphatic, polyheterocycloaliphatic, aromatic or heteroaromaticgroup; and the salts, solvates, hydrates and n-oxides thereof:
 2. Acompound according to claim 1 in which R¹ is an α₄-integrin bindinggroup.
 3. A compound according to claim 1 in which R¹ is a group offormula Ar¹L²Ar²Alk-, where Ar¹ is an optionally substituted aromatic orheteroaromatic group, L² is a linker atom or group, Ar² is an optionallysubstituted phenylene or nitrogen-containing six-membered heteroarylenegroup and Alk is a chain from: —CH₂—CH(R)—, —CH═C(R)—,

where R is a carboxylic acid (—CO₂H) or a derivative or biosterethereof.
 4. A compound according to claim 3 in which Alk is a chainselected from —CH₂—CH(R)—, or


5. A compound according to claim 4 in which R is a carboxylic acid(—CO₂H) group.
 6. A compound according to claim 3 in which Ar² is anoptionally substituted 1,4-phenylene group.
 7. A compound according toclaim 3 in which Ar¹ is an optionally substituted phenyl, monocyclicheteroaromatic or bicycl1c heteroaromatic group.
 8. A compound accordingto claim 7 in which Ar¹ is an optionally substituted pyridyl andpyrimidinyl group. 9 A compound according to claim 8 in which L² is a—CON(R⁸)— group where R⁸ is a hydrogen atom or an optionally substitutedC₁₋₆alkyl group.
 10. A compound according to claim 9 in which R⁸ is ahydrogen atom.
 11. A compound according to claim 7 in which Ar¹ is anoptionally substituted 2,6-naphthyridinyl and 4quinazolinyl groups. 12.A compound according to claim 11 in which L² is an —O— or —N(R⁸)— groupwhere R⁸ is a hydrogen atom or an optionally substituted C₁ 6alkylgroup.
 13. A compound according to claim 12 in which R⁸ is a hydrogenatom.
 14. A compound according to claim 1 in which L¹ is a —N(R⁸)— groupwhere R⁸ is a hydrogen atom or an optionally substituted C₁₋₆alkylgroup.
 15. A compound according to claim 14 in which R⁸ is a hydrogenatom or methyl, ethyl or n-propyl group.
 16. A compound according toclaim 1 in which L¹ is a covalent bond.
 17. A compound according to anyone of claim 1 in which n is the integer 1 and Alk¹ is an optionallysubstituted straight or branched C₁₋₆alkylene chain.
 18. A compoundaccording to claim 17 in which Alk¹ is a —CH₂—, —CH₂CH₂—, —CH₂CH₂CH₂—,—CH(CH₃)CH₂— or —C(CH₃)₂CH₂— chain.
 19. A compound according to claim 18in which R³ is a hydrogen atom.
 20. A compound according to claim 3 offormula (2a):

wherein —W=is —CH= or —N═; R¹⁶ and R¹⁷, which may be the same ordifferent is each a hydrogen atom or group -L³(Alk²)_(t)L⁴(R⁴)_(u) inwhich: L³ is a covalent bond or a linker atom or group; Alk² is analiphatic or heteroaliphatic chain; t is zero or the integer 1; L⁴ is acovalent bond or a linker atom or group; R⁴ is a hydrogen or halogenatom or a group selected from optionally substituted C₁₋₆alkyl or C₃₋₈cycloalkyl, —OR⁵ (where R⁵ is a hydrogen atom, an optionally substituedC₁₋₆alkyl or C₃₋₈ cycloalkyl group], —SR⁵, —NR⁵R⁶ [where R⁶ is as justdefined for R⁵ and may be the same or different], —NO₂, —CN, —CO₂R⁵,—SO₃H, —SOR⁵, —SO₂R⁵, —SO₃R⁵, —OCO₂R⁵, —CONR⁵R⁶, —OCONR⁵R⁶, —CSNR⁵R⁶,—COR⁵, —OCOR⁵, —N(R⁵)COR⁶, —N(R⁵)CSR⁶, —SO₂N(R⁵)(R⁶), —N(R⁵)SO₂R⁶,N(R⁵)CON(R⁶)(R⁷) [where R⁷ is a hydrogen atom, an optionally substitutedC₁₋₆alkyl or C₃₋₈cycloalkyl group], —N(R⁵)CSN(R⁶)(R⁷) or—N(R⁵)SO₂N(R⁶)(R⁷), provided that when t is zero and each of L³ and L⁴is a covalent bond then u is the integer 1 and R⁴ is other than ahydrogen atom; and the salts, solvates, hydrates and N-oxides thereof.21. A compound according to claim 3 of formula (2b)

wherein R¹⁶ is a hydrogen atom or a group -L³(Alk²)_(t)L⁴(R⁴)_(u) inwhich L³ is a covalent bond or a linker atom or group; Alk² is analiphatic or heteroaliphatic chain; t is zero or the integer 1; L⁴ is acovalent bond or a linker atom or group; R⁴ is a hydrogen or halogenatom or a group selected from optionally substituted C₁₋₆alkyl or C₃₋₈cycloalkyl, —OR⁵ [where R⁵ is a hydrogen atom, an optionally substituedC₁₋₆alkyl or C₃₋₈ cycloalkyl group], —SR⁵, —NR⁵R⁶ [where R⁶ is as justdefined for R⁵ and may be the same or different], —NO₂, —CN, —CO₂R⁵,—SO₃H, —SOR⁵, —SO₂R⁵, —SO₃R⁵, —OCO₂R⁵, —CONR⁵R⁶, —OCONR⁵R⁶, —CSNR⁵R⁶,—COR⁵, —OCOR⁵, —N(R⁵)COR⁶, —N(R⁵)CSR⁶, —SO₂N(R⁵)(R⁶), —N(R⁵)SO₂R⁶,N(R⁵)CON(R⁶)(R⁷) [where R⁷ is a hydrogen atom, an optionally substitutedC₁₋₆alkyl or C₃₋₉cycloalkyl group], —N(R⁵)CSN(R⁶)(R⁷) or—N(R⁵)SO₂N(R⁶)(R⁷), provided that when t is zero and each of L³ and L⁴is a covalent bond then u is the integer 1 and R⁴ is other than ahydrogen atom; g is zero or the integer 1, 2, 3 or 4; and the salts,solvates, hydrates and N-oxides thereof.
 22. A compound according toclaim 3 of formula (2c)

wherein R¹⁶ is a hydrogen atom or a group in which L³ is a covalent bondor a linker atom or group; Alk² is an aliphatic or heteroaliphaticchain; t is zero or the integer 1; L⁴ is a covalent bond or a linkeratom or group; R⁴ is a hydrogen or halogen atom or a group selected fromoptionally substituted C₁₋₆alkyl or C₃₋₈ cycloalkyl, —OR⁵ [where R⁵ is ahydrogen atom, an optionally substitued C₁₋₆alkyl or C₃₋₈ cycloalkylgroup], —SR⁵, —NR⁵RO [where R⁶ is as just defined for R⁵ and may be thesame or different], —NO₂, —CN, —CO₂R⁵, —SO₃H, —SOR⁵, —SO₂R⁵, O₃R⁵,—OCO₂R⁵, —CONR⁵R⁶, —OCONR⁵R⁶, —CSNR⁵R⁶, —COR⁵, COR⁵-N(R⁵)COR⁶,—N(R⁵)CSR⁵, —SO₂N(R⁵)(R⁶), —N(R⁵)SO₂R⁶, N(R⁵)CON(R⁶)(R⁷) [where R⁷ is ahydrogen atom, an optionally substituted C₁₋₆alkyl or C₃₋₈cycloalkylgroup], —N(R⁵)CSN(R⁶)(R⁷) or —N(R⁵)SO₂N(R⁶)(R⁷), provided that when t iszero and each of L³ and L⁴ is a covalent bond then u is the integer 1and R⁴ is other than a hydrogen atom; g is zero or the integer 1, 2, 3or 4; and the salts, solvates, hydrates and N-oxides thereof.
 23. Acompound which is:(S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-2-(2-propylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-(3,5-Dichloro4-pyridylcarboxamido)phenyl]-242-t-butyl-3,4-dioxocyclobut-1enyl)amino]propanoic acid;(S)-3-{4[(6,7-Dimethoxy4qunazolinyl)amino]phenyl}2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-([6,7-Dimethoxy-4qunazolinyl)oxy]phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-([6,7-Methoxy4-qunazolinyl]amino)phenyl]-2[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-N,N-dipropylamino-3,4-dioxocyclobut-1enyl)amino]propanoicacid;(S)-3-[4-([2,6-Naphthyridin-1-yl]oxy)phenyl]-2-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-piperidin-1-yl-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(R)-3-{4-(3,5-Dichloro4-pyridylcarboxamido)phenyl)}3-[(2-N,N-diethylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid;(S)-3-[4-([2,6-Naphthyridin-1-yl]oxy)phenyl]-2-[(2-N,N-dipropylamino-3,4-dioxocyclobut-1-enyl)amino]propanoicacid; (S)-3-[4([2,6-Naphthyridin-1-yl]amino)phenyl]-2-[(2-N,-ethyl-N-isopropylamino3,4-dioxocyclobut-1-enyl)amino]propanoic acid;and the salts, solvates, hydrates and N-oxides thereof.
 24. Apharmaceutical composition comprising a compound according to Claim 1together with one or more pharmaceutically acceptable carriers,excipients or diluents.